Methods for the diagnosis of dementia and other neurological disorders

ABSTRACT

The present invention is directed to a method for differentially diagnosing dementia or the risk of dementia in a patient. The method comprises obtaining a sample from the patient; analyzing the sample to obtain quantifying data for one or more than one metabolite marker; comparing the quantifying data for the one or more than one metabolite marker to corresponding data obtained from one or more than one reference sample; and using the comparison to differentially diagnose dementia or the risk of dementia. The method may also assist in assessing dementia or the risk of dementia in a patient. The present invention is also directed to metabolite markers and compounds useful in the present method.

This application is a continuation of U.S. patent application Ser. No.12/280,920, filed Feb. 28, 2007, a national stage application under 35U.S.C. §371 from PCT Application No. PCT/CA2007/000313, filed Feb. 28,2007, which claims the priority benefit of U.S. Provisional ApplicationNos. 60/888,883, filed Feb. 8, 2007, 60/804,779, filed Jun. 14, 2006,and 60/777,290, filed Feb. 28, 2006, each of which is herebyincorporated by reference in its entirety.

FIELD OF INVENTION

The present invention relates to small molecules or metabolites that arefound to have significantly different abundances between clinicallydiagnosed dementia or other neurological disorders, and normal patients.The present invention also relates to methods for diagnosing dementiaand other neurological disorders.

BACKGROUND OF THE INVENTION

The most severe consequence of the aging brain is dementia, which isdefined in the Diagnostic and Statistical Manual of Mental Disorders,4th edition (DSM-IV) as

-   -   “the development of multiple cognitive deficits that included        memory impairment and at least one of the following cognitive        disturbances: aphasia, apraxia, agnosia, or a disturbance in        executive functions. The cognitive impairment must be        sufficiently severe to cause impairment in occupational or        social functioning and must represent a decline from a        previously higher level of functioning.” [1]

The number of elderly people is increasing rapidly within our societyand as a consequence, dementia is growing into a major health problem.In 1991, the Canadian Study of Health and Aging had estimated 25% of thepopulation over the age of 65 had a form of dementia. The study alsoestimated the number of people living with dementia will double andtriple in Canada by 2011 and 2031, respectively [2].

The clinical manifestation of dementia can result from neurodegeneration(e.g. Alzheimer's Disease [AD], dementia with Lewy bodies [DLB] andfrontotemporal lobe dementia [FTLD]), vascular (e.g. multi-infarctdementia) or anoxic event (e.g. cardiac arrest), trauma to the brain(e.g. dementia pugilistica [boxer's dementia]), or exposure to aninfectious (e.g. Creutzfeldt-Jakob Disease) or toxic agent (e.g.alcohol-induced dementia) [3].

AD is the most common cause of dementia, followed by vascular dementia(VaD), DLB and FTLD [4]. The differential diagnosis of the types ofdementia is not straightforward, and is typically based on exclusion ofother disorders [5]. For example, blood chemistry values are measured todetermine if Vitamin B12 deficiency, anemia, infection, venereal diseaseor thyroid disorder may be possible reasons for the dementia symptoms.Various neuroimaging techniques may be employed, such as magneticresonance imaging or computerized tomography scans to determine if thesymptoms may be due to the presence of a tumor, infection or vascularevent [4].

If the dementia symptoms can not be explained by another disorder, adiagnosis of AD, DLB or FTLD is made exclusively based on the clinicalsymptoms (e.g. frequency of falls, rapid onset, presence of visual orauditory hallucinations, etc). It is not until a histopathologicalevaluation of the brain during autopsy is performed that a definitivediagnosis can be obtained [5-7]. A prospective study on the prevalenceof AD in people over the age of 85 indicated that more than half of theindividuals with neuropathological criteria for AD were eithernon-demented or were incorrectly diagnosed with VaD. As well, 35% ofthose individuals diagnosed with AD based on clinical features wereincorrectly diagnosed as the neuropathological evaluation did notsupport that diagnosis [8]. The degree of misdiagnosis is understandablesince the clinical symptoms of the various dementias often overlap andis dependent upon whether the pertinent information is made known to theclinician.

The different types of dementias are based on specific neuropathologicalfeatures. A definitive diagnosis of AD relies on the deposition of twotypes of neuronal protein: tau in the form of intraneuronalneurofibrillary tangles (NFTs) and the accumulation of extracellularβ-amyloid to form senile plaques (SPs). Tau is important for theformation of microtubules in the neuronal axon by binding and promotingpolymerization of tubules. In AD, tau becomes hyperphosphorylatedthereby disrupting its main function. The tau accumulates and formstangles within the axon. The neuron can no longer function and dies. Tauprotein is released into the extracellular space where it can bedetected in the cerebrospinal fluid (CSF) [9]. The formation of SPs,however, is due to the accumulation of a 40 and 42 residue proteinβ-amyloid from amyloid precursor protein (APP) [10]. The formation andsecretion of β-amyloid is closely regulated by homeostasis, butsomething occurs in AD that disrupts homeostasis resulting in theaccumulation of the protein within the brain and disrupting the neuronswithin its vicinity [11-12]. The increased amount of tau and the absenceof β-amyloid in CSF have been proposed as possible diagnostic markersfor AD, but results have not been consistent. The problem may be due tothe presence of NFTs and SPs that increase in number during normal aging[13]. In order for the NFTs and SPs to be diagnostic of AD, they must belocalized together in specific areas of the brain (neocortex and limbicregion) [12]. SPs without NFTs are present in the same area inindividuals with mild cognitive impairment (MCI) and in 27% ofnon-demented individuals greater then 75 years old [13].

A diagnosis of DLB is based on the presence of protein deposits calledalpha-synuclein, which is referred to as Lewy Bodies, within brainstemand cortical neurons [6]. The cognitive deficit corresponds to theamount of Lewy Bodies within the brain.

FTLD is not characterized by a specific neuropathological feature.Typically, areas of the frontal/temporal cortices have neuronal loss,spongiform changes (microvacuolation) and severe astrocytic gliosis. Theclinical symptoms in FTLD are dependent upon where the pathology isfound rather than the type of pathology [7].

Currently, various neuropsychological tests are used to help diagnosedementia. For example, the Alzheimer's Disease Assessment Scale(ADAS)-cognitive subset is used to test the language ability (speech andcomprehension), memory, ability to copy geometric figures andorientation to current time and place. The Folstein's Mini-Mental StateExam (MMSE), which also measures cognitive impairment, is an extensivelyvalidated test of orientation, short and long-term memory, praxis,language and comprehension. While these tests may indicate the level ofcognitive impairment in an individual, they give no indication ofwhether the dementia may be caused by AD or by non-AD dementias.

It is commonly accepted that by the time any symptom is evident in anyof the dementias described, irreversible neuronal loss has occurred[14]. MCI is characterized by a prominent impairment in memory withnormal cognitive functions [15]. MCI is considered a transitional stagebetween normal aging and several types of dementia since a largeproportion of individuals with MCI are later diagnosed with AD, DLB, orFTLD and all individuals with fully developed dementia first exhibitmild dementia symptoms similar to MCI [16].

There is a need to objectively differentiate the types of dementia fromone another. Preferably, such a method would be specific, accurate, andefficient. Clearly, there is a pressing need for differential diagnosisof dementia prior to autopsy.

A biomarker that could detect neuropathological changes prior toclinical symptoms would be of enormous value. A consensus was reached in1999 [17] as to what would be expected in a biomarker in AD:

-   -   1. Detect a fundamental feature of neuropathology    -   2. Diagnostic sensitivity of >80% for detecting AD    -   3. Specificity of >80% for distinguishing other dementias    -   4. Reliable    -   5. Reproducible    -   6. Noninvasive    -   7. Simple to perform    -   8. Inexpensive

The identification of AD-specific biomarkers in human serum would beextremely useful since it would be noninvasive and could be used todetect the presence of AD pathology prior to the manifestation ofclinical symptoms and differentiate those patients who may have adifferent form of dementia but similar clinical symptoms.

SUMMARY OF THE INVENTION

The present invention relates to small molecules or metabolites that arefound to have significantly different abundances between clinicallydiagnosed dementia or other neurological disorders, and normal patients.The present invention also relates to methods for diagnosing dementiaand other neurological disorders.

The present invention provides a method of identifying one or more thanone metabolite marker for differentially diagnosing AD dementia, non-ADdementia, cognitive impairment, or a combination thereof, comprising thesteps of:

-   -   introducing one or more than one sample from one or more than        one patient with clinically diagnosed AD dementia, clinically        diagnosed non-AD dementia, significant cognitive impairment, or        any combination thereof, said sample containing a plurality of        metabolites into a high resolution mass spectrometer obtaining        quantifying data for the metabolites;    -   creating a database of said quantifying data;    -   comparing the identifying and quantifying data from the sample        with corresponding data from a sample from a reference sample;    -   identifying one or more than one metabolite marker that differs        between same sample and said reference sample,

wherein the metabolites metabolite markers are selected from themetabolites listed in Tables 1-7, 10-13, and 18, or any combinationthereof. The method may further comprising selecting a minimal number ofmetabolite markers needed for optimal diagnosis. In a non-limitingexample, the high resolution mass spectrometer is a Fourier TransformIon Cyclotron Resonance Mass Spectrometer (FTMS).

The present invention also provides novel compounds selected from thegroup consisting of the metabolites listed in Tables 7-13. The 15. Themetabolite may be selected from the group consisting ofphosphatidylcholine-related compounds, ethanolamine plasmalogens,endogenous fatty acids, essential fatty acids, lipid oxidationbyproducts, metabolite derivatives of said metabolite classes, and anymetabolite that may contribute in any way to the anabolic/catabolicmetabolism of said metabolite classes.

In one embodiment of the present invention, the compounds may beselected from the group consisting of metabolites with accurate massesmeasured in Daltons of, or substantially equivalent to, 541.3432,569.3687, 699.5198, 723.5195, 723.5197, 751.5555, 803.568, 886.5582,565.3394, 569.369, 801.555, and 857.6186. The metabolites with accuratemasses measured in Daltons of, or substantially equivalent to, a)541.3432, b) 569.3687, c) 699.5198, d) 723.5195, e) 751.5555, and f)803.568 may also be characterized by

-   -   a) an extracted ion chromatogram (EIC) as shown in FIG. 4A, and        an MS/MS spectrum as shown in FIG. 6;    -   b) an EIC as shown in FIG. 4B, and an MS/MS spectrum as shown in        FIG. 7;    -   c) an EIC as shown in FIG. 4C, and an MS/MS spectrum as shown in        FIG. 8;    -   d) an EIC as shown in FIG. 4D, and an MS/MS spectrum as shown in        FIG. 9;    -   e) an EIC as shown in FIG. 4E, and an MS/MS spectrum as shown in        FIG. 10; and    -   f) an EIC as shown in FIG. 4F, and an MS/MS spectrum as shown in        FIG. 11, respectively.

The compounds as described above may also be further characterized bymolecular formula a) C₂₅H₅₁NO₉P, b) C₂₇H₅₅NO₉P, c) C₃₉H₇₄NO₇P, d)C₄₁H₇₄NO₇P, e) C₄₃H₇₈NO₇P, and f) C₄₃H₈₁NO₁₀P, respectively; and/or bythe structures shown in a) FIG. 12; b) FIG. 13; c) FIG. 17; d) FIG. 18;e) FIG. 19; and f) FIG. 14, respectively.

The compounds may also be selected from the group consisting ofmetabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, a) 567.3547, b) 565.3394, c) 805.5832, d)827.57, e) 829.5856, f) 831.5997, and g) 853.5854. These compounds maybe further characterized by molecular formula a) C₂₇H₅₅NO₉P, b)C₂₇H₅₅NO₉P, c) C₄₃H₈₃NO₁₀Y, C₄₅H₈₁NO₁₀Y, e) C₄₅H₈₃NO₁₀P, f) C₄₅H₈₅NO₁₀P,and g) C₄₇H₈₃NO₁₀P, respectively; and/or by the structure shown in a)FIG. 15A; b) FIG. 15B; c) FIG. 15C; d) FIG. 15D; e) FIG. 15E; f) FIG.15F; and g) FIG. 15G, respectively.

The compounds may further be selected from the group consisting ofmetabolites M05 to M24 with accurate masses of, or substantiallyequivalent to those listed in Table 18. Of these compounds, themetabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, a) 701.53591, b) 699.52026, c) 723.52026,d) 747.52026, e) 729.56721, f) 727.55156, g) 779.58286, and h) 775.55156may be further characterized by a MS/MS spectrum as shown in a) FIG. 21;b) FIG. 22; c) FIG. 23; d) FIG. 24; e) FIG. 25; f) FIG. 26; g) FIG. 27;and h) FIG. 28, respectively.

The compounds as described above may also be further characterized bymolecular formula a) C₃₉H₇₆NO₇P, b) C₃₉H₇₄NO₇P, c) C₄₁H₇₄NO₇P, d)C₄₃H₇₄NO₇P, e) C₄₁H₈₀NO₇P, f) C₄₁H₇₈NO₇P, C₄₅H₈₂NO₇P, and h) C₄₅H₇₈NO₇P,respectively and/or by the structure

respectively.

The novel compounds may also be selected from the group consisting ofthe metabolites listed in Table 30. Of these compounds, the metaboliteswith accurate masses measured in Daltons of, or substantially equivalentto, 207.0822, 275.8712, 371.7311, 373.728, 432.1532, 485.5603, 487.6482,562.46, 622.2539, 640.2637, 730.6493, and 742.2972 are of particularinterest.

One or more than one of the compounds of the present invention may beused for the differential diagnosis of dementia.

In another embodiment, the present invention provides a method fordifferentially diagnosing dementia or the risk of dementia in a patient,the method comprising the steps of:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) comparing the quantifying data for said one or more than one        metabolite marker to corresponding data obtained from one or        more than one reference sample; and    -   d) using said comparison to differentially diagnose dementia or        the risk of dementia.

The step of analyzing (step b) may comprise analyzing the sample byliquid chromatography mass spectrometry (LC-MS), or alternatively maycomprise analyzing the sample by liquid chromatography and linear iontrap mass spectrometry when the method is a highthroughput method.

In the method as just described, the one or more than one referencesample is a first reference sample obtained from a non-demented controlindividual. The one or more than one reference sample may also comprisea second reference sample obtained from a patient with clinicallydiagnosed AD-dementia; a third reference sample obtained from a patientwith clinically diagnosed non-AD dementia; and/or a fourth referencesample obtained from a patient suffering from significant cognitiveimpairment.

In one alternative of the method described above, the sample and thereference sample are serum samples, and the one or more than onemetabolite marker is selected from the metabolites listed in Tables 1 to7, or a combination thereof. These metabolite markers may be selectedfrom the group consisting of phosphatidylcholine-related compounds,ethanolamine plasmalogens, endogenous fatty acids, essential fattyacids, lipid oxidation byproducts, metabolite derivatives of saidmetabolite classes, and any metabolite that may contribute in any way tothe anabolic/catabolic metabolism of said metabolite classes.

The one or more than one metabolite marker needed for optimal diagnosismay comprise metabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, 541.3432, 569.3687, 699.5198, 723.5195,723.5197, 751.5555, 803.568, 886.5582, and any combination thereof. Ofthese, the metabolite of accurate masses 699.5198, 723.5195, 723.5197,and 751.555 are ethanolamine plasmalogens and are specifically decreasedin patients with AD dementia; and the metabolite markers of accuratemasses 541.3432, 569.3687, 803.568, and 886.5582 are phosphatidylchoinemetabolites, are decreased in patients with cognitive impairment onADAS-cog, and severity of cognitive impairment correlates to the degreeof decrease.

The one or more than one metabolite marker may be the metabolites withaccurate masses measured in Daltons of, or substantially equivalent to,a) 541.3432, b) 569.3687, c) 699.5198, d) 723.5195, e) 751.5555, and f)803.568. These metabolites may be further characterized by

-   -   a) an extracted ion chromatogram (EIC) as shown in FIG. 4A, and        an MS/MS spectrum as shown in FIG. 6;    -   b) an EIC as shown in FIG. 4B, and an MS/MS spectrum as shown in        FIG. 7;    -   c) an EIC as shown in FIG. 4C, and an MS/MS spectrum as shown in        FIG. 8;    -   d) an EIC as shown in FIG. 4D, and an MS/MS spectrum as shown in        FIG. 9;    -   e) an EIC as shown in FIG. 4E, and an MS/MS spectrum as shown in        FIG. 10; and    -   f) an EIC as shown in FIG. 4F, and an MS/MS spectrum as shown in        FIG. 11,

respectively. The metabolite may also be further characterized bymolecular formula a) C₂₅H₅₁NO₉P, b) C₂₇H₅₅NO₉P, c) C₃₉H₇₄NO₇P, d)C₄₁H₇₄NO₇P, e) C₄₃H₇₈NO₇P, and f) C₄₃H₈₁NO₁₀P, respectively; and/or bythe structure shown in a) FIG. 12; b) FIG. 13; c) FIG. 17; d) FIG. 18;e) FIG. 19; and f) FIG. 14, respectively.

In another alternative of the method described above, the sample and thereference sample may be cerebrospinal fluid (CSF) samples, and the oneor more than one metabolite marker is selected from the metaboliteslisted in Table 13, or a combination thereof. Of particular interest aremetabolite markers needed for optimal diagnosis, which may comprisemetabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, 207.0822, 275.8712, 371.7311, 373.728,432.1532, 485.5603, 487.6482, 562.46, 622.2539, 640.2637, 730.6493,742.2972, and any combination thereof. Of these, metabolite markers207.0822, 432.1532, 562.46, 622.2539, 640.2637, 730.6493, and 742.2972are increased in patients with AD dementia; and metabolite markers275.8712, 371.7311, 373.728, 485.5603, and 487.6482 are decreased inpatients with AD dementia.

In yet another alternative of the method described above, the sample andthe reference sample are serum samples, and the one or more than onemetabolite marker may be selected from metabolites M05 to M24 withaccurate masses of, or substantially equivalent to those listed in Table18. Of these, the one or more than one metabolite marker of particularinterest may comprise metabolites with accurate masses measured inDaltons of, or substantially equivalent to, a) 701.53591, b) 699.52026,c) 723.52026, d) 747.52026, e) 729.56721, f) 727.55156, g) 779.58286,and h) 775.55156, and wherein a decrease in the level of a) to h)indicates AD dementia with a severe cognitive impairment.

The metabolites listed above may be further characterized by a MS/MSspectrum as shown in a) FIG. 21, b) FIG. 22, c) FIG. 23, d) FIG. 24, e)FIG. 25, f) FIG. 26, g) FIG. 27, and h) FIG. 28, respectively. Themetabolites may also be further characterized by molecular formula a)C₃₉H₇₆NO₇P, b) C₃₉H₇₄NO₇P, c) C₄₁H₇₄NO₇P, d) C₄₃H₇₄NO₇P, e) C₄₁H₈₀NO₇P,f) C₄₁H₇₈NO₇P, g) C₄₅H₈₂NO₇P, and h) C₄₅H₇₈NO₇P, respectively; and/or bythe structure

respectively.

In yet another aspect of the present invention, there is provided amethod for assessing dementia or the risk of dementia in a patient, themethod comprising the steps of:

-   -   a) obtaining a serum sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) comparing the quantifying data for said one or more than one        metabolite marker to corresponding data obtained from one or        more than one reference sample; and    -   d) using said comparison to assess dementia or the risk of        dementia.

The step of analyzing (step b) may comprise analyzing the sample byliquid chromatography mass spectrometry (LC-MS), or alternatively maycomprise analyzing the sample by liquid chromatography and linear iontrap mass spectrometry when the method is a highthroughput method.

In the method as just described, the one or more than one referencesample may be a first reference sample obtained from a non-dementedcontrol individual. The one or more than one reference sample may alsofurther comprise a second reference sample obtained from a patient withcognitive impairment as measured by ADAS-cog, and/or a third referencesample obtained from a patient with cognitive impairment as measured byMMSE.

The one or more than one metabolite marker in the method described abovemay be selected from the metabolites listed in Tables 10-12, or acombination thereof. Of particular interest are the one or more than onemetabolite markers is selected from the group consisting of metaboliteswith accurate masses measured in Daltons of, or substantially equivalentto 541.3432, 569.3687, 699.5198, 723.5195, 723.5197, 751.5555, 803.568,886.5582, 565.3394, 569.369, 801.555, 857.6186, and any combinationthereof. Of these, a decrease in the patient sample in metabolitemarkers 699.5198, 723.5195, 723.5197, and 751.555 indicates ADpathology; a decrease in the patient sample in metabolite markers541.3432, 569.3687, 803.568, and 886.5582 indicates cognitive impairmenton ADAS-cog; and a decrease in the patient sample in metabolite markers565.3394, 569.369, 801.555, and 857.6186 indicates cognitive impairmenton MMSE.

In yet another embodiment of the present invention, a method is providedfor differentially diagnosing dementia or the risk of dementia in apatient, the method comprising the steps of:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) obtaining a ratio for each of the one or more than one        metabolite marker to an internal control metabolite;    -   d) comparing each ratio of said one or more than one metabolite        marker to the internal control metabolite to corresponding data        obtained from one or more than one reference sample; and    -   e) using said comparison to differentially diagnose dementia or        the risk of dementia.

The step of analyzing (step b) may comprise analyzing the sample byliquid chromatography mass spectrometry (LC-MS), or alternatively maycomprise analyzing the sample by liquid chromatography and linear iontrap mass spectrometry when the method is a highthroughput method.

In the method as just described, the one or more than one referencesample may be a first reference sample obtained from a non-dementedcontrol individual. The one or more than one reference sample mayfurther comprise a second reference sample obtained from a patient withclinically diagnosed AD-dementia; a third reference sample obtained froma patient with clinically diagnosed non-AD dementia; and/or a fourthreference sample obtained from a patient suffering from significantcognitive impairment.

In one aspect of the method described above, the sample and thereference sample are serum samples, and the one or more than onemetabolite marker is selected from metabolites M05 to M24 with accuratemasses of, or substantially equivalent to those listed in Table 18. Ofparticular interest is the one or more than one metabolite markercomprising metabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, a) 701.53591, b) 699.52026, c) 723.52026,d) 747.52026, e) 729.56721, f) 727.55156, g) 779.58286, and h)775.55156, and the internal control metabolite comprising the metabolitewith accurate mass measured in Daltons of, or substantially equivalentto, 719.54648. When these metabolites and internal control metaboliteare used, a decrease in the ratio of metabolite to the internal controlmetabolite indicates AD dementia with a severe cognitive impairment.

The metabolites described above may be further characterized by a MS/MSspectrum as shown in a) FIG. 21, b) FIG. 22, c) FIG. 23, d) FIG. 24, e)FIG. 25, f) FIG. 26, g) FIG. 27, and h) FIG. 28, respectively. Thesemetabolites may also be further characterized by molecular formula a)C₃₉H₇₆NO₇P, b) C₃₉H₇₄NO₇P, c) C₄₁H₇₄NO₇P, d) C₄₃H₇₄NO₇P, e) C₄₁H₈₀NO₇P,C₄₁H₇₈NO₇P, g) C₄₅H₈₂NO₇P, and h) C₄₅H₇₈NO₇P, respectively, and theinternal control metabolite may be characterized by molecular formulaC₃₉H₇₈NO₈P; and/or by the structure

respectively, and the internal control metabolite may be furthercharacterized by the structure

In yet another embodiment of the present invention, there is provided amethod for evaluating the efficacy of a therapy for treating dementia ina patient, comprising:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) comparing said quantifying data to corresponding data        obtained from one or more than one reference sample; and    -   d) using said comparison to determine whether the therapy is        improving the demented state of the patient.

The step of analyzing (step b) may comprise analyzing the sample byliquid chromatography mass spectrometry (LC-MS), or alternatively maycomprise analyzing the sample by liquid chromatography and linear iontrap mass spectrometry when the method is a highthroughput method.

In the method as just described, the one or more than one referencesample may be a plurality of samples obtained from a non-dementedcontrol individuals; a plurality of samples obtained from a clinicallydiagnosed AD patient; one or more than one pre-therapy baseline sampleobtained from the patient; or any combination thereof.

In one aspect of the above method, the sample and the reference sampleare serum samples, and the one or more than one metabolite marker isselected from the metabolites listed in Tables 1 to 7, or a combinationthereof. These metabolite marker markers needed for optimal diagnosismay be selected from the group consisting of phosphatidylcholine-relatedcompounds, ethanolamine plasmalogens, endogenous fatty acids, essentialfatty acids, lipid oxidation byproducts, metabolite derivatives of saidmetabolite classes, and any metabolite that may contribute in any way tothe anabolic/catabolic metabolism of said metabolite classes. Ofparticular interest are the metabolites with accurate masses measured inDaltons of, or substantially equivalent to, 541.3432, 569.3687,699.5198, 723.5195, 723.5197, 751.5555, 803.568, 886.5582.

In another aspect, the sample and the reference sample are cerebrospinalfluid (CSF) samples, and the one or more than one metabolite marker isselected from the metabolites listed in Table 13, or a combinationthereof. Of particular interest are the metabolites with accurate massesmeasured in Daltons of, or substantially equivalent to, 207.0822,275.8712, 371.7311, 373.728, 432.1532, 485.5603, 487.6482, 562.46,622.2539, 640.2637, 730.6493, 742.2972.

In a third aspect, the sample and the reference sample are serumsamples, and the one or more than one metabolite marker may be selectedfrom metabolites M05 to M24 with accurate masses of, or substantiallyequivalent to those listed in Table 18. Of these metabolites, themetabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, 701.53591, 699.52026, 723.52026, 747.52026,729.56721, 727.55156, 779.58286, and 775.55156 may be of particularinterest.

The present invention also provides a method for evaluating the efficacyof a therapy for treating dementia in a patient, comprising:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) obtaining a ratio for each of the one or more than one        metabolite marker to an internal control metabolite;    -   d) comparing each ratio of said one or more than one metabolite        marker to the internal control metabolite to corresponding data        obtained from one or more than one reference sample; and    -   e) using said comparison to determine whether the therapy is        improving the demented state of the patient.

The step of analyzing (step b) may comprise analyzing the sample byliquid chromatography mass spectrometry (LC-MS), or alternatively maycomprise analyzing the sample by liquid chromatography and linear iontrap mass spectrometry when the method is a highthroughput method.

In the method as just described, the one or more than one referencesample may be a plurality of samples obtained from a non-dementedcontrol individuals; a plurality of samples obtained from a clinicallydiagnosed AD patient; one or more than one pre-therapy baseline sampleobtained from the patient; or any combination thereof.

In the method as described above, the sample and said reference sampleare serum samples, and the one or more than one metabolite marker may beselected from metabolites M05 to M24 with accurate masses of, orsubstantially equivalent to those listed in Table 18. Of particularinterest are the metabolites with accurate masses measured in Daltonsof, or substantially equivalent to, 701.53591, 699.52026, 723.52026,747.52026, 729.56721, 727.55156, 779.58286, and 775.55156, and theinternal control metabolite with accurate mass measured in Daltons of,or substantially equivalent to, 719.54648.

The methods of the present invention, including HTS assays, can be usedfor the following, wherein the specific “health-state” in thisapplication refers to, but is not limited to dementia:

1. identifying small-molecule metabolite biomarkers which candiscriminate between multiple health-states using any biological sampletaken from an individual,

2. specifically diagnosing a health-state using metabolites identifiedin serum, plasma, whole blood, serum, CSF, and/or other tissue biopsy asdescribed in this application,

3. selecting the minimal number of metabolite features required foroptimal diagnostic assay performance statistics using supervisedstatistical methods such as those mentioned in this application,

4. identifying structural characteristics of biomarker metabolitesselected from non-targeted metabolomic analysis using LC-MS/MS, MSn andNMR,

5. developing a high-throughput LC-MS/MS method for assaying selectedmetabolite levels in serum,

6. diagnosing a given health-state, or risk for development of ahealth-state by determining the levels of any combination of metabolitefeatures disclosed from the FTMS analysis patient serum, using anymethod including but not limited to mass spectrometry, NMR, UVdetection, ELISA (enzyme-linked immunosorbant assay), chemical reaction,image analysis, or other.

The impact of the present invention on the diagnosis of dementia wouldbe tremendous, as literally everyone could be screened longitudinallythroughout their lifetime to assess risk. Given that the performancecharacteristics of the test of the present invention are representativefor the general population, this test alone may be superior to any othercurrently available screening method, as it may have the potential todetect disease progression prior to the emergence of clinical symptoms.

This summary of the invention does not necessarily describe all featuresof the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features of the invention will become more apparent fromthe following description in which reference is made to the appendeddrawings wherein:

FIG. 1 shows a mean signal-to-noise +/−SEM of the AD serum 8 biomarkerpanel for each different clinical group (AD with significant cognitiveimpairment, Non-AD dementia, and AD with no significant cognitiveimpairment) relative to non-demented controls.

FIG. 2 shows a mean signal-to-noise +/−SEM of the AD serum 8 biomarkerpanel for two clinical groups with a significant cognitive impairment(AD and Non-AD dementia).

FIG. 3 shows a mean signal-to-noise +/−SEM of the AD CSF 12 biomarkerpanel for two clinical groups with a significant cognitive impairment(AD and Non-AD dementia).

FIG. 4 shows Q-Star extracted ion chromatograms (EIC) for themetabolites 541.3432 (A), 569.3687 (B), 699.5198 (C), 723.5195 (D),751.5555 (E), and 803.568 (F). Top panel, 8 samples from non-dementedsubjects, bottom panel, 8 samples from clinically-diagnosed AD subjects.

FIG. 5 shows averaged AD biomarker intensities of the 8 AD and 8non-demented controls samples from FTMS and Q-Star Analysis.

FIG. 6 shows MS/MS spectra for metabolite 541.3432 with CE voltage −50V.

FIG. 7 shows MS/MS spectra for metabolite 569.3687 with CE voltage −50V.

FIG. 8 shows MS/MS spectra for metabolite 699.5198 with CE voltage −50V.

FIG. 9 shows MS/MS spectra for metabolite 723.5195 with CE voltage −50V.

FIG. 10 shows MS/MS spectra for metabolite 751.5555 with CE voltage−50V.

FIG. 11 shows MS/MS spectra for metabolite 803.568 with CE voltage −50V.

FIG. 12 shows structural determination of ADAS-cog serum biomarker541.3432.

FIG. 13 shows structural determination of ADAS-cog serum biomarker569.3687.

FIG. 14 shows structural determination of ADAS-cog serum biomarker803.568.

FIG. 15 shows putative structures of additional serum biomarkers.A—metabolite with mass of 567.3547; B—metabolite with mass of 565.3394;C—metabolite with mass of 805.5832; D—metabolite with mass of 827.57;E—metabolite with mass of 829.5856; F—metabolite with mass of 531.5997;and G—metabolite with mass of 853.5854.

FIG. 16 shows the fragments obtained for the MS/MS analysis of the751.5555 metabolite, along with its proposed structure.

FIG. 17 shows the fragments obtained for the MS/MS analysis of the699.5198 metabolite, along with its proposed structure.

FIG. 18 shows the fragments obtained for the MS/MS analysis of the723.5195 metabolite, along with its proposed structure.

FIG. 19 shows the LC-MS and MS/MS analysis of the 751.5555 metabolite(18:0/20:4 EtnPls). Panel A1 is an extracted ion chromatogram (EIC) ofparent ion 750 (M—H—) of a pure standard; panel A2 is MS/MS spectra ofparent ion M/Z 750 @ retention time 4.8-5.0 minutes. Panel B1 is the EICof parent ion 750 from a cognitively normal subject; panel B2 is theMS/MS spectra of parent ion M/Z 750 @ 4.8-5.0 min. Panel C1 is the EICof parent ion 750 from an AD subject; and panel C2 is the MS/MS spectraof parent ion M/Z 750@ 4.8-5.0 min.

FIG. 20 shows the general structure of ethanolamine phospholipids, aswell as the naming convention used herein.

FIG. 21 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 16:0/18:1 (M15) in human serum.

FIG. 22 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 16:0/18:2 (M16) in human serum.

FIG. 23 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 16:0/20:4 (M17) in human serum.

FIG. 24 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 16:0/22:6 (M19) in human serum.

FIG. 25 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 18:0/18:1 (M20) in human serum.

FIG. 26 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 18:0/18:2 (M21) in human serum.

FIG. 27 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 18:0/20:4 (M23) in human serum.

FIG. 28 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 18:0/22:6 (M24) in human serum.

FIG. 29 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 18:1/18:2 and Plasmanyl 16:0/20:4 (M07)in human serum.

FIG. 30 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of EtnPls 20:0/20:4 and EtnPls 18:0/22:4 (M23) inhuman serum.

FIG. 31 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panel) of Plasmanyl 18:0/20:4 (M12) and Plasmanyl16:0/22:4 (M08) in human serum.

FIG. 32 is an extracted ion chromatogram (upper panel) and MS/MSspectrum (lower panels) of EtnPls 18:1/20:4, EtnPls 16:0/22:5, Plasmanyl16:0/22:6 (M09) in human serum.

FIG. 33 shows a Q-Trap flow injection analysis standard curve of EtnPls16:0/22:6 (M19) in healthy human serum.

FIG. 34 shows the effect of dementia severity and SDAT pathology onserum EtnPl levels (male and female subjects combined). (A) Mono anddi-unsaturated EtnPls and saturated PtdEt internal control. (B)Polyunsaturated EtnPls and free DHA (22:6). EtnPls abbreviations: (fattyacid carbons:double bonds, not including the vinyl ether double bond)and position on glycerol backbone (sn-1/sn-2). D16:0/18:0 representsdiacylglycerophosphatidylethanolamine with palmitic acid (16:0) at sn-1and stearic acid (18:0) at sn-2; 22:6 represents free DHA. Values areexpressed as mean+SEM (n=19-112).

FIG. 35 shows serum DHA-EtnPls (Log(2) EtnPls 16:0/22:6 (M19) to PtdEt16:0/18:0 (M01) ratio) distributions in subjects with different levelsof dementia severity (male and female subjects combined).

FIG. 36 is a comparison of theoretical distributions of AD pathology (A)compiled from references 5-8 and experimentally determined distributionsof serum 22:6-containing EtnPls (Log(2) EtnPls 16:0/22:6 (M19) to PtdEt16:0/18:0 (M01) ratio) (B) in cognitively normal and demented subjects.Arrow indicates positive diagnosis of AD.

FIG. 37 is a linear regression analysis of disease severity (ADAS-cog)and serum 22:6-containing EtnPls (EtnPls 16:0/22:6 (M19) to PtdEt16:0/18:0 (M01) ratio) levels in 256 AD subjects. X=predicted initiationof EtnPls depletion. Values are expressed as mean±SEM (n=66-112).Clinical progression assumes 7.5 ADAS-cog points/year.

FIG. 38 shows serum 22:6-containing EtnPls (EtnPls 16:0/22:6 (M19) toPtdEt 16:0/18:0 (M01) ratio) levels in AD, Cognitive Normal, and generalpopulation subjects. (A) Mean±SEM (n=68-256). (B) Log(2) distributions.

FIG. 39 shows the distribution of serum white and gray matter EtnPlscores in males and females.

DETAILED DESCRIPTION

The present invention relates to small molecules or metabolites that arefound to have significantly different abundances between clinicallydiagnosed dementia or other neurological disorders, and normal patients.The present invention also relates to methods for diagnosing dementiaand other neurological disorders.

The present invention provides novel methods for discovering,validating, and implementing a metabolite markers for one or morediseases or particular health-states. In one embodiment of the presentinvention, there is provided a method for identifying specificbiomarkers for differentially diagnosing AD dementia, non-AD dementia,cognitive impairment, or a combination thereof, comprising the steps of:introducing one or more than one sample from one or more than onepatient with clinically diagnosed AD dementia, clinically diagnosednon-AD dementia, or significant cognitive impairment, said samplecontaining a plurality of metabolites into a high resolution massspectrometer (for example, and without wishing to be limiting, a FourierTransform Ion Cyclotron Resonance Mass Spectrometer (FTMS)); obtaining,identifying and quantifying data for the metabolites; creating adatabase of said identifying and quantifying data; comparing theidentifying and quantifying data from the sample with corresponding datafrom a sample from a non-demented normal patient; identifying one ormore than one metabolites that differ. The metabolite markers identifiedusing the method of the present invention may include the metaboliteslisted in Tables 1-7, 10-13, and 18. The method may further compriseselecting the minimal number of metabolite markers needed for optimaldiagnosis.

In order to determine the biochemical markers of a given health-state ina particular population, a group of patients representative of thehealth state (i.e. a particular disease) and/or a group of “normal”counterparts are required. Biological samples taken from the patients inthe particular health-state category can then be compared to the samesamples taken from the normal population as well as to patients insimilar health-state category in the hopes of identifying biochemicaldifferences between the two groups, by analyzing the biochemicalspresent in the samples using FTMS and/or LC-MS.

The method for the discovery of metabolite markers as described abovemay be done using non-targeted metabolomic strategies or methods.Multiple non-targeted metabolomics strategies have been described in thescientific literature including NMR [18], GC-MS [19-21], LC-MS, and FTMSstrategies [18, 22-24]. The metabolic profiling strategy employed forthe discovery of differentially expressed metabolites in the presentinvention was the non-targeted FTMS strategy by Phenomenome Discoveries[21, 24-27; see also US Published Application No. 2004-0029120 A1,Canadian Application No. 2,298,181, and WO 0157518]. Non-targetedanalysis involves the measurement of as many molecules in a sample aspossible, without any prior knowledge or selection of components priorto the analysis. Therefore, the potential for non-targeted analysis todiscover novel metabolite biomarkers is high versus targeted methods,which detect a predefined list of molecules. The present invention usesa non-targeted method to identify metabolite components in serum samplesthat differ between clinically diagnosed AD individuals and non ADindividuals. The same technology was used to identify metabolitecomponents that differ between clinically diagnosed AD individuals withdementia from clinically diagnosed non-AD individuals with dementia inCSF samples.

However, a person skilled in the art would recognize that othermetabolite profiling strategies could be used to discover some or all ofthe differentially regulated metabolites disclosed in the presentinvention and that the metabolites described herein, however discoveredor measured, represent unique chemical entities that are independent ofthe analytical technology that may be used to detect and measure them.

The present invention also provides a method for differentiallydiagnosing dementia or the risk of dementia in a patient, the methodcomprising the steps of:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) comparing the quantifying data for said one or more than one        metabolite marker to corresponding data obtained from one or        more than one reference sample; and    -   d) using said comparison to differentially diagnose dementia or        the risk of dementia.        The step of analyzing the sample (steb b) may comprise analyzing        the sample using a mass spectrometer (MS). For example, and        without wishing to be limiting, such mass spectrometer could be        of the FTMS, orbitrap, time of flight (TOF) or quadrupole types.        Alternatively, the mass spectrometer could be equipped with an        additional pre-detector mass filter. For example, and without        wishing to be limiting such instruments are commonly referred to        as quadrupole-FTMS (Q-FTMS), quadrupole-TOF (Q-TOF) or triple        quadrupole (TQ or QQQ). In addition, the mass spectrometer could        be operated in either the parent ion detection mode (MS) or in        MSn mode, where n>=2. MSn refers to the situation where the        parent ion is fragmented by collision induced dissociation (CID)        or other fragmentation procedures to create fragment ions, and        then one or more than one of said fragments are detected by the        mass spectrometer. Such fragments can then be further fragmented        to create further fragments. Alternatively, the sample could be        introduced into the mass spectrometer using a liquid or gas        chromatographic system or by direct injection.

By the term “differential diagnosis” or “differentially diagnosing”, itis meant that various aspects of a disease state may be distinguishedfrom one another. In particular, the present invention allows fordifferential diagnosis a various states of dementia; for example andwithout wishing to be limiting, the present invention may providedifferential diagnosis of AD dementia, non-AD dementia, cognitiveimpairment, or a combination thereof.

The diagnosis of or the exclusion of any types of neurological disordersis contemplated by the present invention, using all or a subset of themetabolites disclosed herein. The term “dementia” is used herein as abroad term indicating both cognitive impairment as well as pathologiescausing cognitive impairment. Dementia may be caused by a number ofneurological disorders. “AD dementia” as used herein refers to dementiacaused by Alzheimer's disease (AD, which may also be referred to hereinas “SDAT”); types of “non-AD dementia” include, but are not limited to,dementia with Lewy bodies (DLB), frontotemporal lobe dementia (FTD),vascular induced dementia (e.g. multi-infarct dementia), anoxic eventinduced dementia (e.g. cardiac arrest), trauma to the brain induceddementia (e.g. dementia pugilistica [boxer's dementia]), dementiaresulting from exposure to an infectious (e.g. Creutzfeldt-JakobDisease) or toxic agent (e.g. alcohol-induced dementia), Autism,Multiple Sclerosis, Parkinson's Disease, Bipolar Disorder, Ischemia,Huntington's Chorea, Major Depressive Disorder, Closed Head Injury,Hydrocephalus, Amnesia, Anxiety Disorder, Traumatic Brain Injury,Obsessive Compulsive Disorder, Schizophrenia, Mental Retardation, and/orEpilepsy. Of particular interest are AD dementia, and FTD and DLB non-ADdementias.

Cognitive impairment can be assessed by any method known in the art. Forexample, and without wishing to be limiting, the Alzheimer's DiseaseAssessment Scale (ADAS)-cognitive subset may be used. Thisneuropsychological test is used to test the language ability (speech andcomprehension), memory, ability to copy geometric figures andorientation to current time and place. Errors on the test are recordedresulting in a reverse score impairment (i.e., the higher the score onADAS, the greater the cognitive impairment). A score of 0-15 isconsidered normal, 16-47 is considered mild-moderate impairment and ascore of 48-70 is considered moderate-severe impairment [28]. Anotherneuropsychological test, Folstein's Mini-Mental State Exam (MMSE), whichmeasures cognitive impairment, may be used. The MMSE is widely used andis an extensively validated test of orientation, short and long-termmemory, praxis, language and comprehension. A person skilled in the artwould recognize that additional neuropsychological assessment thatmeasure aspects of the same cognitive deficit, such as, but notexclusive to, the Blessed Roth Dementia Rating Scale, the 7-MinuteScreen, Wechsler Memory Scale (WMS), Halstead-Reitan Battery, ReyAuditory Verbal Learning Test, California Verbal Learning Test, BuschkeSelective Reminding Test, Boston Naming Test, Clinical Evaluation ofLanguage Functioning, Peabody Picture Vocabulary Tests, Mattis DementiaRating Scale, Memory Assessment Scale, Tests of Memory and Learning,Wide Range Assessment of Memory and Learning, can also be used.

In addition, a person skilled in the art would recognize that anyimaging technique that has the potential to show a cognitive impairmentor structural change, such as, but not exclusive to, structural magneticresonance imaging (MRI), positron emission tomography (PET),computerized tomography (CT), functional magnetic resonance imaging(fMRI), electroencephalography (EEG), single positron emissiontomography (SPECT), event related potentials, magnetoencephalography,multi-modal imaging, would be measuring the structural/regional brainareas that are responsible for that cognitive deficit and AD pathology,and therefore, would be related to the metabolites disclosed in thisinvention.

In accordance with the present invention, any type of biological samplethat originates from anywhere within the body, for example but notlimited to, blood (serum/plasma), CSF, urine, stool, breath, saliva, orbiopsy of any solid tissue including tumor, adjacent normal, smooth andskeletal muscle, adipose tissue, liver, skin, hair, brain, kidney,pancreas, lung, colon, stomach, or other may be used. Of particularinterest are samples that are serum or CSF. While the term “serum” isused herein, those skilled in the art will recognize that plasma orwhole blood or a sub-fraction of whole blood may also be used. CSF maybe obtained by a lumbar puncture requiring a local anesthetic.

In a non-limiting example, when a blood sample is drawn from a patientthere are several ways in which the sample can be processed. The rangeof processing can be as little as none (i.e. frozen whole blood) or ascomplex as the isolation of a particular cell type. The most common androutine procedures involve the preparation of either serum or plasmafrom whole blood. All blood sample processing methods, includingspotting of blood samples onto solid-phase supports, such as filterpaper or other immobile materials, are also contemplated by the presentinvention.

In another non-limiting example, CSF samples may be collected using alumbar puncture procedure; a local anesthetic is applied to the lowerback. A needle is then inserted into the numbed skin between the L4 andL5 vertebrae until it pierces the subdural space. The CSF may becollected into sterile tubes.

For example, but not considered to be limiting in any manner, whileobtaining a CSF sample may result in more discomfort for the patientthan taking a blood sample, a CSF assay used after a positive result ona AD-specific serum test, a differential diagnosis between AD and non-ADhas a higher degree of confirmation.

Without wishing to be limiting in any manner, the processed blood, serumor CSF sample described above may then be further processed to make itcompatible with the methodical analysis technique to be employed in thedetection and measurement of the metabolites contained within theprocessed serum or CSF sample. The types of processing can range from aslittle as no further processing to as complex as differential extractionand chemical derivatization. Extraction methods could includesonication, soxhlet extraction, microwave assisted extraction (MAE),supercritical fluid extraction (SFE), accelerated solvent extraction(ASE), pressurized liquid extraction (PLE), pressurized hot waterextraction (PHWE) and/or surfactant assisted extraction (PHWE) in commonsolvents such as methanol, ethanol, mixtures of alcohols and water, ororganic solvents such as ethyl acetate or hexane. A method of particularinterest for extracting metabolites for FTMS non-targeted analysis is toperform a liquid/liquid extraction whereby non-polar metabolitesdissolve in an organic solvent and polar metabolites dissolve in anaqueous solvent.

The extracted samples may be analyzed using any suitable method know inthe art. For example, and without wishing to be limiting in any manner,extracts of biological samples are amenable to analysis on essentiallyany mass spectrometry platform, either by direct injection or followingchromatographic separation. Typical mass spectrometers are comprised ofa source which ionizes molecules within the sample, and a detector fordetecting the ionized molecules or fragments of molecules. Non-limitingexamples of common sources include electron impact, electrosprayionization (ESI), atmospheric pressure chemical ionization (APCI),atmospheric pressure photo ionization (APPI), matrix assisted laserdesorption ionization (MALDI), surface enhanced laser desorptionionization (SELDI), and derivations thereof. Common mass separation anddetection systems can include quadrupole, quadrupole ion trap, linearion trap, time-of-flight (TOF), magnetic sector, ion cyclotron (FTMS),Orbitrap, and derivations and combinations thereof. The advantage ofFTMS over other MS-based platforms is its high resolving capability thatallows for the separation of metabolites differing by only hundredths ofa Dalton, many which would be missed by lower resolution instruments.

By the term “metabolite”, it is meant specific small molecules, thelevels or intensities of which are measured in a sample, and that may beused as markers to diagnose a disease state. These small molecules mayalso be referred to herein as “metabolite marker”, “metabolitecomponent”, “biomarker”, or “biochemical marker”.

The metabolites are generally characterized by their accurate mass, asmeasured by mass spectormetry technique used in the above method. Theaccurate mass may also be referred to as “accurate neutral mass” or“neutral mass”. The accurate mass of a metabolite is given herein inDaltons (Da), or a mass substantially equivalent thereto. By“substantially equivalent thereto”, it is meant that a +/−5 ppmdifference in the accurate mass would indicate the same metabolite, aswould be recognized by a person of skill in the art. The accurate massis given as the mass of the neutral metabolite. As would be recognizedby a person of skill in the art, the ionization of the metabolites,which occurs during analysis of the sample, the metabolite will causeeither a loss or gain of one or more hydrogen atoms and a loss or gainof an electron. This changes the accurate mass to the “ionized mass”,which differs from the accurate mass by the mass of hydrogens andelectrons lost or gained during ionization. Unless otherwise specified,the accurate neutral mass will be referred to herein.

Similarly, when a metabolite is described by its molecular formula orstructure, the molecular formula or structure of the neutral metabolitewill be given. Naturally, the molecular formula or structure of theionized metabolite will differ from the neutral molecular formula orstructure by the number of hydrogens lost or gained during ionization.

Data is collected during analysis and quantifying data for one or morethan one metabolite is obtained. “Quantifying data” is obtained bymeasuring the levels or intensities of specific metabolites present in asample.

The quantifying data is compared to corresponding data from one or morethan one reference sample. The “reference sample” is any suitablereference sample for the particular disease state. For example, andwithout wishing to be limiting in any manner, in the present inventionthe reference sample may be a sample from a non-demented controlindividual, i.e., a person not suffering from AD dementia, non-ADdementia or cognitive impairment (also referred to herein as a “‘normal’counterpart”); the reference sample may also be a sample obtained from apatient with clinically diagnosed with AD, a patient with clinicallydiagnosed non-AD dementia, or a patient diagnosed with significantcognitive impairment. As would be understood by a person of skill in theart, more than one reference sample may be used for comparison to thequantifying data. For example and without wishing to be limiting, theone or more than one reference sample may be a first reference sampleobtained from a non-demented control individual. The one or more thanone reference sample may further include a second reference sampleobtained from a patient with clinically diagnosed AD-dementia, a thirdreference sample obtained from a patient with clinically diagnosednon-AD dementia, a fourth reference sample obtained from a patientsuffering from significant cognitive impairment, or any combinationthereof.

The present invention also provides novel compounds, identified usingthe methods of the present invention. The novel compounds may be used asmetabolite markers in the differential diagnosis of dementia, asdescribed above.

In one embodiment, the compounds may be selected from the metaboliteslisted in Tables 1 to 7, or a combination thereof. These metaboliteswere identified in serum samples, and may be phosphatidylcholine-relatedcompounds, ethanolamine plasmalogens, endogenous fatty acids, essentialfatty acids, lipid oxidation byproducts, metabolite derivatives of saidmetabolite classes, and any metabolite that may contribute in any way tothe anabolic/catabolic metabolism of said metabolite classes.

An optimal panel of compounds may be indentified from those metaboliteslisted in Tables 1 to 7. For example and without wishing to be limiting,the metabolite markers may be metabolites with accurate masses measuredin Daltons of, or substantially equivalent to, 541.3432, 569.3687,699.5198, 723.5195, 723.5197, 751.5555, 803.568, 886.5582. Themetabolites of accurate masses 699.5198, 723.5195, 723.5197, and 751.555have presently been identified as ethanolamine plasmalogens, and arespecifically decreased in patients with AD dementia. The metabolitemarkers of accurate masses 541.3432, 569.3687, 803.568, and 886.5582have presently been identified as phosphatidylcholine relatedmetabolites, and are decreased in patients with cognitive impairment onADAS-cog, and severity of cognitive impairment correlates to the degreeof decrease.

The metabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, a) 541.3432, b) 569.3687, c) 699.5198, d)723.5195, e) 751.5555, f) 803.568, can be further characterized by

-   -   a) an extracted ion chromatogram (EIC) as shown in FIG. 4A, and        an MS/MS spectrum as shown in FIG. 6; a molecular formula of        C₂₅H₅₁NO₉P; and/or the structure shown in FIG. 12;    -   b) an EIC as shown in FIG. 4B, and an MS/MS spectrum as shown in        FIG. 7; a molecular formula of C₂₇H₅₅NO₉P; and/or the structure        shown in FIG. 13;    -   c) an EIC as shown in FIG. 4C, and an MS/MS spectrum as shown in        FIG. 8; a molecular formula of C₃₉H₇₄NO₇P; and/or the structure        shown in FIG. 17;    -   d) an EIC as shown in FIG. 4D, and an MS/MS spectrum as shown in        FIG. 9; a molecular formula of C₄₁H₇₄NO₇P; and/or the structure        shown in FIG. 18;    -   e) an EIC as shown in FIG. 4E, and an MS/MS spectrum as shown in        FIG. 10; a molecular formula of C₄₃H₇₈NO₇P; and/or the structure        shown in FIG. 19;    -   f) an EIC as shown in FIG. 4F, and an MS/MS spectrum as shown in        FIG. 11; a molecular formula of C₄₃H₈₁NO₁₀P; and/or the        structure shown in FIG. 14,    -   respectively.

It is presently shown that the ethanolamine plasmalogen metabolites(neutral masses 699.5198, 723.5195, 751.5555) and thephosphatidylcholine metabolites (neutral masses 699.5198, 723.5195,751.5555) are decreased in the serum of AD subjects exhibitingsignificant cognitive impairment. This is the first report ofserum-based changes in these metabolites associated with AD anddementia. It is further shown that the decrease in the disclosed serumphospatidylcholine related metabolites occurs in all patients exhibitinga significant cognitive impairment as measured by the ADAS-cogregardless of AD status, and that the degree of decrease correlates withthe severity of the cognitive impairment. However, the observed decreasein disclosed ethanolamine plasmalogens is independent of cognitiveimpairment, occurs specifically in subjects with AD and is therefore atrue diagnostic of AD.

Ethanolamine plasmalogens are a type of ethanolamine phospholipid.Ethanolamine phospholipids can be further differentiated based on theirsn-1 configurations (either acyl, ether, or vinyl ether). The sn-2position is typically acyl and the sn-3 position contains thephosphoethanolamine moiety. Therefore, the three classes are describedas either diacyl (also referred to herein as PtdEt), alkyl-acyl (alsoreferred to herein as plasmanyl) or alkenyl-acyl (also referred toherein as EtnPl or plasmenyl). Various basic structures of ethanolaminephospholipids are shown in FIG. 20, along with the standard namingconvention used herein.

A decrease in the disclosed ethanolamine plasmalogens may represent theinitial or early stages AD, and can be detected non-invasively in livingsubjects by measuring serum levels of specific ethanolamineplasmalogens. Similarly, cognitive impairment can be quantitatednon-invasively by measuring the serum levels of specificphosphatidylcholine metabolites.

Other metabolites have also been identified. For example, themetabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, a) 567.3547, b) 565.3394, c) 805.5832, d)827.57, e) 829.5856, f) 831.5997, and g) 853.5854, which may be furthercharacterized by

-   -   a) the molecular formula C₂₇H₅₅NO₉P; and/or the structure shown        in FIG. 15A    -   b) the molecular formula C₂₇H₅₅NO₉P; and/or the structure shown        in FIG. 15B    -   c) the molecular formula C₄₃H₈₃NO₁₀P; and/or the structure shown        in FIG. 15C;    -   d) the molecular formula C₄₅H₈₁NO₁₀P; and/or the structure shown        in FIG. 15D;    -   e) the molecular formula C₄₅H₈₃NO₁₀P; and/or the structure shown        in FIG. 15E;    -   f) the molecular formula C₄₅H₈₅NO₁₀P; and/or the structure shown        in FIG. 15F;    -   g) the molecular formula C₄₇H₈₃NO₁₀P; and/or the structure shown        in FIG. 15G,    -   respectively.

Based on the identification of metabolites specific to AD dementia(accurate masses 699.5198, 723.5195, 723.5197, 751.555) as ethanolamineplasmalogens, other ethanolamine phospholipid metabolite markers wereidentified. These are metabolites M05 to M24 as listed and characterized(accurate mass, name/composition, molecular formula) in Table 18. Thestructure of the metabolite can be deduced based on the metabolite nameas indicated in Table 18 and the nomenclature, as indicated in FIG. 20.

Of the compounds listed in Table 18, those of particular interestinclude metabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, a) 701.53591, b) 699.52026, c) 723.52026,d) 747.52026, e) 729.56721, f) 727.55156, g) 779.58286, and h)775.55156, which can be further characterized by

-   -   a) a MS/MS spectrum as shown in FIG. 21; molecular formula        C₂₇H₅₅NO₉P; and/or the structure

-   -   b) a MS/MS spectrum as shown in FIG. 22; molecular formula        C₃₉H₇₄NO₇P; and/or the structure

-   -   c) a MS/MS spectrum as shown in FIG. 23; molecular formula        C₄₁H₇₄NO₇P; and/or the structure

-   -   d) a MS/MS spectrum as shown in FIG. 24; molecular formula        C₄₃H₇₄NO₇P; and/or the structure

-   -   e) a MS/MS spectrum as shown in FIG. 25; molecular formula        C₄₁H₈₀NO₇P; and/or the structure

-   -   f) a MS/MS spectrum as shown in FIG. 26; molecular formula        C₄₁H₇₈NO₇P; and/or the structure

-   -   g) a MS/MS spectrum as shown in FIG. 27; molecular formula        C₄₅H₈₂NO₇P; and/or the structure

-   -   h) a MS/MS spectrum as shown in FIG. 28; molecular formula        C₄₅H₇₈NO₇P; and/or the structure

-   -   respectively.

In another embodiment of the present invention, the compounds may beselected from the metabolites listed in Table 13, or a combinationthereof. These metabolites were identified in CSF samples. Of particularinterest are the metabolites with accurate masses measured in Daltonsof, or substantially equivalent to, 207.0822, 275.8712, 371.7311,373.728, 432.1532, 485.5603, 487.6482, 562.46, 622.2539, 640.2637,730.6493, 742.2972. When used to diagnose dementia, the metabolitemarkers 207.0822, 432.1532, 562.46, 622.2539, 640.2637, 730.6493, and742.2972 are increased in patients with AD dementia; and metabolitemarkers 275.8712, 371.7311, 373.728, 485.5603, and 487.6482 aredecreased in patients with AD dementia.

In a further method of the present invention, a method for assessingdementia or the risk of dementia in a patient is described. The methodcomprises the steps of:

-   -   a) obtaining a serum sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) comparing the quantifying data for said one or more than one        metabolite marker to corresponding data obtained from one or        more than one reference sample; and    -   d) using said comparison to assess dementia or the risk of        dementia.

The step of analyzing the sample (steb b)) may comprise analyzing thesample by liquid chromatography mass spectrometry (LC-MS).Alternatively, the step of analyzing the sample (step b)) may compriseanalyzing the sample by linear ion trap mass spectrometry followed byliquid chromatograph, when the method is a highthroughput method.

The one or more than one reference sample may include a first referencesample obtained from a non-demented control individual, a secondreference sample obtained from a patient with cognitive impairment asmeasured by ADAS-cog, a third reference sample obtained from a patientwith cognitive impairment as measured by MMSE, or a combination of oneor more of these.

Without wishing to be limiting in any manner, the one or more than onemetabolite marker used to asses dementia or the risk of dementia may beselected from the metabolites listed in Tables 10-12, or a combinationthereof. Of particular interest are metabolites with accurate massesmeasured in Daltons of, or substantially equivalent to 541.3432,569.3687, 699.5198, 723.5195, 723.5197, 751.5555, 803.568, 886.5582,565.3394, 569.369, 801.555, 857.6186. A decrease in the patient samplein metabolite markers 699.5198, 723.5195, 723.5197, and 751.555indicates AD pathology; a decrease in the patient sample in metabolitemarkers 541.3432, 569.3687, 803.568, and 886.5582 indicates cognitiveimpairment on ADAS-cog; and 565.3394, 569.369, 801.555, and 857.6186indicates cognitive impairment on MMSE.

In yet another embodiment of the present invention, there is provided amethod for differentially diagnosing dementia or the risk of dementia ina patient. The method comprising the steps of:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) obtaining a ratio for each of the one or more than one        metabolite marker to an internal control metabolite;    -   d) comparing each ratio of said one or more than one metabolite        marker to the internal control metabolite to corresponding data        obtained from one or more than one reference sample; and    -   e) using said comparison to differentially diagnose dementia or        the risk of dementia.

The step of analyzing the sample (steb b) may comprise analyzing thesample using a mass spectrometer (MS). For example, and without wishingto be limiting, such mass spectrometer could be of the FTMS, orbitrap,time of flight (TOF) or quadrupole types. Alternatively, the massspectrometer could be equipped with an additional pre-detector massfilter. For example, and without wishing to be limiting such instrumentsare commonly referred to as quadrupole-FTMS (Q-FTMS), quadrupole-TOF(Q-TOF) or triple quadrupole (TQ or QQQ). In addition, the massspectrometer could be operated in either the parent ion detection mode(MS) or in MSn mode, where n>=2. MSn refers to the situation where theparent ion is fragmented by collision induced dissociation (CID) orother fragmentation procedures to create fragment ions, and then one ormore than one of said fragments are detected by the mass spectrometer.Such fragments can then be further fragmented to create furtherfragments. Alternatively, the sample could be introduced into the massspectrometer using a liquid or gas chromatographic system or by directinjection.

In the method as just described above, the one or more than onereference sample may be a first reference sample obtained from anon-demented control individual. The one or more than one referencesample may further include a second reference sample obtained from apatient with clinically diagnosed AD-dementia, a third reference sampleobtained from a patient with clinically diagnosed non-AD dementia, afourth reference sample obtained from a patient suffering fromsignificant cognitive impairment, or any combination thereof.

In the method as described above, the sample and reference sample may beserum samples. The one or more than one metabolite marker may beselected from the metabolites as listed and characterized (accuratemass, name/composition, molecular formula) in Table 18. The “internalcontrol metabolite” refers to an endogenous metabolite naturally presentin the patient. Any suitable endogenous metabolite that does not varyover the disease states can be used as the internal control metabolite.For example, and without wishing to be limiting, the internal controlmetabolite may be phosphatidylethanolamine 16:0/18:0 (PtdEt 16:0/18:0,M01), as shown in Table 18; this internal control metabolite has amolecular formula of C₁₉H₇₈NO₈P, and a structure characterized as

Use of the ratio of the metabolite marker to the internal controlmetabolite offers measurement that are more stable and reproducible thanmeasurement of absolute levels of the metabolite marker. As the internalcontrol metabolite is naturally present in all samples and does notappear to vary significantly over disease states, the sample-to-samplevariability (due to handling, extraction, etc) is minimized.

Of the compounds listed in Table 18, those of particular interest in theabove method include metabolites with accurate masses measured inDaltons of, or substantially equivalent to, a) 701.53591, b) 699.52026,c) 723.52026, d) 747.52026, e) 729.56721, f) 727.55156, g) 779.58286,and h) 775.55156. A decrease in the ratio of a) to h) to the internalcontrol metabolite indicates AD dementia with a severe cognitiveimpairment. These metabolites can be further characterized by

-   -   a) a MS/MS spectrum as shown in FIG. 21; molecular formula        C₂₇H₅₅NO₉P; and/or the structure

-   -   b) a MS/MS spectrum as shown in FIG. 22; molecular formula        C₃₉H₇₄NO₇P; and/or the structure

-   -   c) a MS/MS spectrum as shown in FIG. 23; molecular formula        C₄₁H₇₄NO₇P; and/or the structure

-   -   d) a MS/MS spectrum as shown in FIG. 24; molecular formula        C₄₃H₇₄NO₇P; and/or the structure

-   -   e) a MS/MS spectrum as shown in FIG. 25; molecular formula        C₄₁H₈₀NO₇P; and/or the structure

-   -   f) a MS/MS spectrum as shown in FIG. 26; molecular formula        C₄₁H₇₈NO₇P; and/or the structure

-   -   g) a MS/MS spectrum as shown in FIG. 27; molecular formula        C₄₅H₈₂NO₇P; and/or the structure

-   -   h) a MS/MS spectrum as shown in FIG. 28; molecular formula        C₄₅H₇₈NO₇P; and/or the structure

-   -   respectively.

In yet another embodiment of the present invention, there is provided amethod for evaluating the efficacy of a therapy for treating dementia ina patient, comprising:

-   -   a) obtaining a sample from said patient;    -   b) analyzing said sample to obtain quantifying data for one or        more than one metabolite marker;    -   c) comparing said quantifying data to corresponding data        obtained from one or more than one reference sample; and    -   d) using said comparison to determine whether the therapy is        improving the demented state of the patient.

Optionally, after the step of analyzing (step b), a ratio for each ofthe one or more than one metabolite marker to an internal controlmetabolite may be obtained. In this case, each ratio of said one or morethan one metabolite marker to the internal control metabolite tocorresponding data obtained from one or more than one reference sampleis compared to evaluate the efficacy of the therapy.

The step of analyzing (step b) may comprise analyzing the sample byliquid chromatography mass spectrometry (LC-MS), or alternatively maycomprise analyzing the sample by liquid chromatography and linear iontrap mass spectrometry when the method is a highthroughput method.

By the term “therapy”, it is meant any suitable course of therapy thatmay improve the health state or demented state of the patient beingevaluated. When evaluating the efficacy of the therapy, the effect ofthe particular therapy in improving or degrading the health state of thepatient will be measured. In doing so, a person of skill in the artwould be capable of determining whether the therapy is effective fortreating the demented state.

In the methods as described, the one or more than one reference samplemay be any suitable reference sample. For example, and without wishingto be limiting in any manner, the reference sample may be a plurality ofsamples obtained from non-demented control individuals; a plurality ofsamples obtained from clinically diagnosed AD patients; one or more thanone pre-therapy baseline sample obtained from the patient; or anycombination thereof. A pre-therapy baseline sample from the patient isparticularly useful, as the variation in metabolites will then bespecific to the patient.

The sample and the reference sample may be serum samples. In this case,the one or more than one metabolite marker could be selected from themetabolites listed in Tables 1 to 7, or a combination thereof, forexample, metabolite markers with accurate masses measured in Daltons of,or substantially equivalent to, 541.3432, 569.3687, 699.5198, 723.5195,723.5197, 751.5555, 803.568, 886.5582. Alternatively, the metabolitemarkers may be selected from metabolites M05 to M24 with accurate massesof, or substantially equivalent to those listed in Table 18, forexample, metabolites with accurate masses measured in Daltons of, orsubstantially equivalent to, 701.53591, 699.52026, 723.52026, 747.52026,729.56721, 727.55156, 779.58286, and 775.55156. Metabolites M05-M24could also be used when a ratio is obtained between the metabolites andthe internal control metabolite; the internal metabolite could be, forexample, metabolite M01, as described in Table 18.

The sample and the reference sample may also be cerebrospinal fluid(CSF) samples. In this case, the one or more than one metabolite markercould be selected from the metabolites listed in Table 13, or acombination thereof; for example, metabolites with accurate massesmeasured in Daltons of, or substantially equivalent to, 207.0822,275.8712, 371.7311, 373.728, 432.1532, 485.5603, 487.6482, 562.46,622.2539, 640.2637, 730.6493, 742.2972.

The identified metabolites can be readily measured systemically. Thispoint is of fundamental importance, since the majority of researchpertaining to AD and other neurological disorders has ignored theperipheral systems. The ability to measure neurodegenerative processeswithin a blood sample is of substantial value in the diagnosis ofdementia. With respect to the specific ethanolamine plasmalogenmetabolites of the present invention, these are a valid biochemicalmarker of AD pathology since this molecular species' content does notchange in Parkinson's disease, a disease which is often accompanied bydementia [29]. Furthermore, the specificity of the plasmalogenmetabolites to AD indicates that its levels in serum could be readilymeasured longitudinally throughout the lifetime of an individual toassess the risk or for early detection of the disease prior to theemergence of clinical symptoms.

The present invention also provides high throughput methods fordifferential diagnosis of AD dementia and non-AD dementia states. Themethod may involve fragmentation of the parent molecule; in anon-limiting example, this may be accomplished by a Q-Trap™ system.Detection of the metabolites may be performed using one of various assayplatforms, including colorimetric chemical assays (UV, or otherwavelength), antibody-based enzyme-linked immunosorbant assays (ELISAs),chip-based and polymerase-chain reaction for nucleic acid detectionassays, bead-based nucleic-acid detection methods, dipstick chemicalassays or other chemical reaction, image analysis such as magneticresonance imaging (MRI), positron emission tomography (PET) scan,computerized tomography (CT) scan, nuclear magnetic resonance (NMR), andvarious mass spectrometry-based systems.

A high-throughput method for determining the levels of the metabolitesin a person's blood and comparing the levels to levels in a normal“reference” population can lead to a prediction of whether the personhas AD or not. This can be carried out in several ways. One way is touse a prediction algorithm to classify the test sample, as previouslydescribed, which would output a percentage probability for having AD. Apredictive approach would work independently of the assay method, aslong as the intensities of the metabolites could be measured. Anothermethod could simply be based on setting a threshold intensity level fromthe mass spectrometer, and determining whether a person's profile isabove or below the threshold which would indicate their AD status.Alternatively, and without wishing to be limiting in any manner, apreferred method is a truly quantitative assay could be performed todetermine the molar concentration of the six metabolites in thenon-demented normal and AD population. An absolute thresholdconcentration could then be determined for AD-positivity. In a clinicalsetting, this would mean that if the measured levels of the metabolites,or combinations of the metabolites, were below a certain concentration,there would be an associated probability that the individual is positivefor AD. Therefore, the optimal diagnostic test could comprise a methodof measuring the intensities of the metabolites in serum, and analgorithm for taking the intensity values and outputting a predictedprobability for having AD as well as for being healthy (i.e.,AD-negative).

The methods and identified biomarkers of the present invention, based onsmall molecules or metabolites in a sample, fulfills the criteriaidentified in 1999 for an ideal screening test [82], as development ofassays capable of detecting specific metabolites is relatively simpleand cost effective per assay. The test is minimally invasive and isindicative of cognitive impairment and of AD pathology. Translation ofthe method into a clinical assay compatible with current clinicalchemistry laboratory hardware is commercially acceptable and effective.Furthermore, the method of the present invention does not require highlytrained personnel to perform and interpret the test.

The present invention will be further illustrated in the followingexamples.

Example 1 Identification of Differentially Expressed Metabolites

Differentially expressed metabolites were identified in clinicallydiagnosed AD with and without significant cognitive impairment,clinically diagnosed non-AD and non-demented controls.

Clinical Samples.

For the AD serum diagnostic assay described, samples were obtained fromrepresentative populations of non-demented healthy individuals and ofclinically diagnosed AD and non-AD dementia patients. The biochemicalmarkers of AD described in the invention were derived from the analysisof 75 serum samples from patients clinically diagnosed with probable AD(43 patients with significant cognitive impairment, 32 with no cognitiveimpairment), serum samples from 30 patients with clinically diagnosednon-AD dementia, and 31 serum samples from non-demented controls.Samples in the three groups were from a diverse population ofindividuals, ranging in age, ethnicity, weight, occupation, anddisplaying varying non-dementia-related health-states. All samples weresingle time-point collections. Cognitive impairment of the patients wasalso assessed using the Alzheimer's Disease Assessment Scale(ADAS)-cognitive subset.

For the AD CSF diagnostic assay described, samples were obtained from agroup of patients that represented clinically diagnosed AD with dementiaand non-AD patients with dementia. The biochemical markers of ADdescribed in this invention were derived from the analysis of 6 CSFsamples from clinically diagnosed AD patients with dementia and 5 CSFsamples from clinically diagnosed non-AD patients with dementia.

Samples in both groups were from a diverse population of individuals,ranging in age, ethnicity, weight, occupation, and displaying varyingnon-dementia-related health-states. All samples were single time-pointcollections. The metabolites contained within the 136 serum samples and11 CSF samples used in this application were separated into polar andnon-polar extracts through sonication and vigorous mixing (vortexmixing).

Mass Spectrometry Analysis.

Analysis of serum extracts collected from 136 individuals (75 clinicallydiagnosed AD, 30 clinically diagnosed non-AD, and 31 non-dementedhealthy controls) and 11 CSF extracts (6 clinically-diagnosed AD and 5clinically diagnosed non-AD patients) was performed by direct injectioninto a FTMS and ionization by either ESI or atmospheric pressurechemical ionization (APCI) in both positive and negative modes. Sampleextracts were diluted either three or six-fold in methanol:0.1% (v/v)ammonium hydroxide (50:50, v/v) for negative ionization modes, or inmethanol:0.1% (v/v) formic acid (50:50, v/v) for positive ionizationmodes. For APCI, sample extracts were directly injected withoutdiluting. All analyses were performed on a Bruker Daltonics APEX IIIFourier transform ion cyclotron resonance mass spectrometer equippedwith a 7.0 T actively shielded superconducting magnet (Bruker Daltonics,Billerica, Mass.). Samples were directly injected using electrosprayionization (ESI) and/or APCI at a flow rate of 1200 μL per hour. Iontransfer/detection parameters were optimized using a standard mix ofserine, tetra-alanine, reserpine, Hewlett-Packard tuning mix and theadrenocorticotrophic hormone fragment 4-10. In addition, the instrumentconditions were tuned to optimize ion intensity and broad-bandaccumulation over the mass range of 100-1000 amu according to theinstrument manufacturer's recommendations. A mixture of theabovementioned standards was used to internally calibrate each samplespectrum for mass accuracy over the acquisition range of 100-1000 amu.

In total, six separate analyses comprising combinations of extracts andionization modes were obtained for each sample:

Aqueous Extract

-   -   1. Positive ESI (analysis mode 1101)    -   2. Negative ESI (analysis mode 1102)

Organic Extract

-   -   3. Positive ESI (analysis mode 1201)    -   4. Negative ESI (analysis mode 1202)    -   5. Positive APCI (analysis mode 1203)    -   6. Negative APCI (analysis mode 1204)

Mass Spectrometry Data Processing.

Using a linear least-squares regression line, mass axis values werecalibrated such that each internal standard mass peak had a mass errorof <1 p.p.m. compared with its theoretical mass. Using XMASS softwarefrom Bruker Daltonics Inc., data file sizes of 1 megaword were acquiredand zero-filled to 2 megawords. A sinm data transformation was performedprior to Fourier transform and magnitude calculations. The mass spectrafrom each analysis were integrated, creating a peak list that containedthe accurate mass and absolute intensity of each peak. Compounds in therange of 100-2000 m/z were analyzed. In order to compare and summarizedata across different ionization modes and polarities, all detected masspeaks were converted to their corresponding neutral masses assuminghydrogen adduct formation. A self-generated two-dimensional (mass vs.sample intensity) array was then created using DISCOVAmetrics™ software(Phenomenome Discoveries Inc., Saskatoon, SK, Canada). The data frommultiple files were integrated and this combined file was then processedto determine the unique masses. The average of each unique mass wasdetermined, representing the y axis. This value represents the averageof all of the detected accurate masses that were statisticallydetermined to be equivalent. Considering that the mass accuracy of theinstrument for the calibration standards is approximately 1 ppm, aperson skilled in the art will recognize that these average masses mayinclude individual masses that fall within +/−5 ppm of this averagemass. A column was created for each file that was originally selected tobe analyzed, representing the x axis. The intensity for each mass foundin each of the files selected was then filled into its representativex,y coordinate. Coordinates that did not contain an intensity value wereleft blank. Once in the array, the data were further processed,visualized and interpreted, and putative chemical identities wereassigned. Each of the spectra were then peak picked to obtain the massand intensity of all metabolites detected. These data from all of themodes were then merged to create one data file per sample. The data fromall 136 samples was then merged and aligned to create a two-dimensionalmetabolite array in which each sample is represented by a column andeach unique metabolite is represented by a single row. In the cellcorresponding to a given metabolite sample combination, the intensity ofthe metabolite in that sample is displayed. When the data is representedin this format, metabolites showing differences between groups ofsamples were determined. The same procedure was utilized to combine the11 CSF samples in a two-dimensional metabolite array.

A. Serum Biomarkers

A student's T-test was used to select for metabolites which differedsignificantly between the following different clinical groups in serum.Metabolites that were less than p<0.05 were considered significant.

A1—Clinically diagnosed AD patients (n=75) vs. non-demented controls(n=31). This comparison yielded 262 metabolites (see Table 1).

A2—Clinically diagnosed AD patients with a significant cognitiveimpairment (n=32) vs. non-demented controls (n=31). This comparisonyielded 292 metabolites (see Table 2).

A3—Clinically diagnosed AD patients with a significant cognitiveimpairment (n=32) vs. clinically diagnosed non-AD patients with asignificant cognitive impairment (n=30); this comparison yielded 118metabolites markers (see Table 3).

A4—Clinically diagnosed AD patients with significant cognitiveimpairment (n=32) vs. clinically diagnosed AD patients withoutsignificant cognitive impairment (n=43). This comparison yielded 97metabolites markers (see Table 4).

A5—Clinically diagnosed non-AD patients (n=30) vs. non-demented controls(n=31); this comparison yielded 199 metabolites markers (see Table 5).

A6—Clinically diagnosed AD patients with mild cognitive impairment(n=43) vs. non-demented controls (n=31). This comparison yielded 136metabolites (see Table 6).

A7—Patients with significant cognitive impairment (n=42) and patientswith a mild cognitive impairment (n=43). This comparison yielded 81metabolites (Table 7).

Tables 1-7 show biochemical markers whose concentrations or amounts inserum are significantly different (p<0.05) between the testedpopulations and therefore have potential diagnostic utility foridentifying each of the aforesaid populations. The features aredescribed by their accurate mass and analysis mode, which together aresufficient to provide the putative molecular formulas and chemicalcharacteristics (such as polarity and putative functional groups) foreach metabolite.

From the initial lists of several hundred possible metabolites, it wasdetermined that a combination of 8 metabolites fulfills the criteria fora serum dementia test: the combination can differentiate AD dementiafrom non-AD dementia, the early stages of AD and healthy controls. Thebest combination of 8 metabolites included the metabolites with neutralmasses (measured in Daltons) 541.3432, 569.3687, 699.5198, 723.5195,723.5197, 751.5555, 803.568, 886.5582. Although these are the actualmasses, a person skilled in the art of this technology would recognizethat +/−5 ppm difference would indicate the same metabolite.

In analyzing the present results, a person of skill in the art wouldunderstand that the following clinical groups are of interest: non-ADwith significant cognitive impairment, AD with significant cognitiveimpairment, AD without significant cognitive impairment and non-dementedcontrols. Bar graphs representing the mean+/−SEM of the 8 biomarkers forthe four different clinical groups are shown in FIG. 1. Relative tocontrol, non-demented individuals, the three non-control states can bedescribed as follows:

1. Non-AD with significant cognitive impairment vs. control:

-   -   a. Biomarker 541.3432—decreased    -   b. Biomarker 569.3687—decreased    -   c. Biomarker 699.5198—no difference    -   d. Biomarker 723.5195—no difference    -   e. Biomarker 723.5197—no difference    -   f. Biomarker 751.5555—no difference    -   g. Biomarker 803.568—decreased    -   h. Biomarker 886.5582—decreased

2. Clinically diagnosed AD with significant cognitive impairment vs.control

-   -   a. Biomarker 541.3432—decreased    -   b. Biomarker 569.3687—decreased    -   c. Biomarker 699.5198—decreased    -   d. Biomarker 723.5195—decreased    -   e. Biomarker 723.5197—decreased    -   f. Biomarker 751.5555—decreased    -   g. Biomarker 803.568—decreased    -   h. Biomarker 886.5582—decreased    -   3. Clinically diagnosed AD without significant cognitive        impairment vs. control    -   a. Biomarker 541.3432—decreased    -   b. Biomarker 569.3687—no difference    -   c. Biomarker 699.5198—decreased    -   d. Biomarker 723.5195—decreased    -   e. Biomarker 723.5197—decreased    -   f. Biomarker 751.5555—decreased    -   g. Biomarker 803.568—no difference    -   h. Biomarker 886.5582—no difference

In each of the three non-control cases described above, a unique subsetof markers was decreased.

Bar graphs representing the mean+/−SEM of the 8 biomarkers for the twodifferent clinical groups with a significant cognitive impairment areshown in FIG. 2. Relative to non-AD dementia with significant cognitiveimpairment, AD patients with significant cognitive impairment can bedescribed as:

-   -   a. Biomarker 541.3432—no difference    -   b. Biomarker 569.3687—no difference    -   c. Biomarker 699.5198—decreased    -   d. Biomarker 723.5195—decreased    -   e. Biomarker 723.5197—decreased    -   f. Biomarker 751.5555—decreased    -   g. Biomarker 803.568—no difference    -   h. Biomarker 886.5582—no difference

The results of this invention show a clear distinction between the serumof individuals with clinically diagnosed AD WITH a significant cognitiveimpairment, individuals with clinically diagnosed AD WITHOUT asignificant cognitive impairment (this could be early stage AD),individuals with non-AD dementia WITH a significant cognitiveimpairment, and non-demented controls. These findings are capable ofidentifying and distinguishing the different types of dementia from oneanother and from the early stages of cognitive impairment as describedin this application. From the above results, it can be further concludedthat the metabolite markers with masses 699.5198, 723.5195, 723.5997,751.5555 are specific for AD pathology; while markers with masses of541.3432, 569.3687, 803.568, 886.5582 are specific for cognitiveimpaired based on ADAS-cog testing.

A second neuropsychological test, Folstein's Mini-Mental State Exam(MMSE), which measures cognitive impairment, was applied to all 136patients. The MMSE is widely used and is an extensively validated testof orientation, short and long-term memory, praxis, language andcomprehension. In the clinically diagnosed AD patients that had nosignificant cognitive impairment (n=43), 15 of those patients had ascore on MMSE that would indicate normal cognition (MMSE≧28), whereasthe remaining 28 patients had MMSE scores that indicated a mildimpairment (score 18-23, n=11) or severe cognitive impairment (score9-17, n=17). A F-test was used to select for metabolites which differedsignificantly between the MMSE scores (normal, mild or severe cognitiveimpairment) for 43 clinically diagnosed AD patients with no significantcognitive impairment on the ADAS-cog test (p<0.05). 23 metabolites metthis criterion (shown in Table 8). These are all features which differstatistically between the two populations and therefore have potentialdiagnostic utility. The features are described by their accurate massand analysis mode, which together are sufficient to provide the putativemolecular formulas and chemical characteristics (such as polarity andputative functional groups) of each metabolite.

An optimal subset of 4 metabolites, all of which were observed todecrease, from the 23 metabolites was selected using PrincipalComponents Analysis (PCA). The 4 metabolites able to produce thegreatest separation between the groups were 565.3394, 569.369, 801.555,857.6186. The metabolites are indicated by asterisks on Table 8 andrepresent a 4-metabolite biomarker panel associated with cognitiveimpairment on MMSE. The fact that a second set of metabolites wereassociated cognitive impairment suggests that the MMSE must be specificto one or several other cognitive states that the ADAS-cog is notspecifically measuring.

Therefore, a total of three 4-biomarker panels can be applied to the 136patients to classify them into one of 8 categories which willsimultaneously indicate the presence of AD pathology (biomarkers699.5198, 723.5195, 723.5997, 751.5555), cognitive impaired on ADAS-cog(541.3432, 569.3687, 803.568, 886.5582) and cognitive impaired on MMSE(565.3394, 569.369, 801.555, 857.6186). Using a 0/1 binary model, eachpatient can be labeled using a 3 digit code from “000” indicating nocognitive impairment and no AD pathology to “111” indicating both MMSEand ADAS-cog impairment and AD pathology. Table 9 indicates theseparation of the patient samples into the 8 categories.

The three 4-biomarker panels were applied individually to the metabolitearray and the patients that showed the best separation on the PCA plotwere selected. These patients were selected because they represented thebest discriminator between the 3 different groups [AD (n=20) vs. non-ADpathology (n=20), high ADAS score (n=20) vs. low ADAS score (n=12),impaired cognition on the MMSE score (n=20) vs. normal cognition on theMMSE score (n=20)]. A student's t-test was performed between thedifferent clinical groups (p<0.05). The 116 metabolites that met thep-value criteria for AD vs. non-AD pathology are listed in Table 10.Table 11 lists the 124 metabolites that met the p-value criteria forhigh ADAS score vs. low ADAS score, and Table 12 contains the list of344 metabolites that met the p-value criteria for impaired score on MMSEand normal cognition on MMSE.

Both the ADAS-cog and MMSE neuropsychological tests measure cognitiveerrors related to praxis, orientation, memory and language ability.Therefore, it would be reasonable to suggest biomarkers associated withADAS-cog score and/or MMSE are related to the ability to conceive of,organize and initiate unfamiliar sequences, the awareness of one's selfand environment, as well as memory and language ability. As such, thesebiomarkers are not exclusive to cognitive impairment associated withdementia; rather any condition that results in any type of praxis,orientation, memory and/or language deficit would show a similarreduction within a biological sample.

The sample set (136 individuals) used for this discovery was nottrivial, and was comprised of individuals of various ethnic andgeographical backgrounds, and of varying age and health status.Therefore, there is sound reason to expect that the findings arerepresentative of the general dementia population.

B. CSF Biomarkers.

A student's T-test was used to select for metabolites which differbetween the clinically diagnosed AD patients and clinically diagnosednon-AD patients in CSF samples (p<0.05). 42 metabolites met thiscriterion (shown in Table 13). These metabolites differed statisticallybetween the two populations and therefore have potential diagnosticutility. The metabolites are described by their accurate mass andanalysis mode, which together are sufficient to provide the putativemolecular formulas and chemical characteristics (such as polarity andputative functional groups) of each metabolite.

An optimal subset of 12 metabolites from the 42 metabolites describedabove was selected. These metabolites had the greatest statisticaldifference between the two groups (p<0.01). Metabolites were excluded ifthey were not detected in at least 60% of the samples in each group (4/6clinically diagnosed AD and 3/5 clinically diagnosed non-AD). The panelcomprises masses 207.0822, 275.8712, 371.7311, 373.728, 432.1532,485.5603, 487.6482, 562.46, 622.2539, 640.2637, 730.6493, 742.2972.Although these are the actual masses, a person skilled in the art ofthis technology would recognize that an +/−5 ppm difference wouldindicate the same metabolite.

The 12 biomarker panel was tested using 5 CSF samples from undiagnosedpatients. The only information available on the samples was thesubject's age, gender, and whether an individual had a cognitivedeficit. If the 12 biomarker panel was correct, the subject could bediagnosed as having AD dementia, non-AD dementia, or normal. From the 5CSF samples provided by undiagnosed patients, 1 was diagnosed withnon-AD dementia, 2 with AD dementia, and 2 as normal. The two normalsubjects did not have a cognitive impairment as indicated by the MiniMental State Examination (MMSE) score. Therefore, using a 12 metabolitefeature set it was possible to both diagnose AD and non-AD dementia.

Bar graphs representing the mean+/−SEM of the 12 biomarkers for the twodifferent clinical groups are shown in FIG. 3. Relative to non-ADdementia with significant cognitive impairment, AD patients with asignificant cognitive impairment can be described as:

-   -   a. Biomarker 207.0822—increased    -   b. Biomarker 275.8712—decreased    -   c. Biomarker 371.7311—decreased    -   d. Biomarker 373.728—decreased    -   e. Biomarker 432.1532—increased    -   f. Biomarker 485.5603—decreased    -   g. Biomarker 487.6482—decreased    -   h. Biomarker 562.46—increased    -   i. Biomarker 622.2539—increased    -   j. Biomarker 640.2637—increased    -   k. Biomarker 730.6493—increased    -   l. Biomarker 742.2972—increased

Based on these results, a clear distinction was made between the CSF ofclinically diagnosed non-AD and AD patients. Therefore, such findingsare capable of identifying and distinguishing AD dementia from non-ADdementia and can form the basis of a dementia diagnostic test in CSF asdescribed in this application. It is expected that the finding arerepresentative of the general dementia population.

Although a non-targeted FTMS-based platform was used in theidentification and selection of the optimal metabolites in serum andCSF, other methods of subsequently detecting the molecules, includingother MS-based platforms, ELISAs, colorimetric assays, etc can used todetect the molecules.

Example 2 Independent Method Confirmation of Discovered Metabolites

A. Serum Biomarkers

An independent mass spectrometry method was used to verify the intensitydifferences between non-demented normal and clinically-diagnosed ADserums of the eight diagnostic metabolites discovered using the FTMSmethod. Eight representative clinically-diagnosed AD sample extracts andeight representative non-demented control sample extracts were analyzedby LC-MS using an HP 1100 high-performance liquid chromatographyinterfaced to an ABI Q-Star mass spectrometer.

Aqueous fractions from five clinically-diagnosed AD and fivenon-demented control sample extracts were evaporated under nitrogen gasand reconstituted in 100 uL of methanol:water:formic acid (5:94.9:0.1).Five μL of the reconstituted sample was subjected to HPLC (AgilentTechnologies) (HP 1100 with Metasil AQ 3u, 100×2 mm column) for fullscan and 10 μL for MS/MS at a flow rate of 0.2 ml/min.

Eluate from the HPLC was analyzed using an ABI Q-Star XL massspectrometer fitted with a Turboion spray ion (ESI) source in negativemode. The scan type in full scan mode was time-of-flight (TOF) with anaccumulation time of 1.0000 seconds, mass range between 50 and 1500 Da,and duration time of 70 min. Source parameters were as follows: Ionsource gas 1 (GS1) 55; Ion source gas 2 (GS2) 90; Curtain gas (CUR) 40;Nebulizer Current (NC) 0; Temperature 450° C.; Declustering Potential(DP)-55; Focusing Potential (FP)-265; Declustering Potential 2 (DP2)-15.In MS/MS mode, scan type was product ion, accumulation time was 1.0000seconds, scan range between 50 and 1000 Da and duration time 70 min. Allsource parameters are the same as above, with a collision energy of (CE)of −50 V and collision gas (CAD, nitrogen) of 5 psi.

Six of the eight metabolite masses previously discovered on the FTMSwere verified on the ABI Q-Star mass spectrometer. The metabolites withthe accurate masses of 723.5195 and 723.5197 were determined to be thesame metabolite, and the metabolite with accurate mass of 886.5582 wasnot detected. Therefore, only six metabolites (699.5198, 723.5195,751.5555, 541.3432, 569.3687, 803.568) were used for the remaininganalyses.

The extracted ion chromatograms (EICs) for the six biomarkers are shownin FIG. 4 The top panel shows the eight non-demented control EICs, andthe bottom panel of each shows the eight clinically-diagnosed AD EICs.The sensitivity of the Q-star is superior to the FTMS, resulting in agreater magnitude in intensity difference between the non-dementedcontrol subjects and clinically diagnosed AD population for the selectedbiomarkers. FIG. 5 shows the average raw intensity of the six biomarkersof the eight non-demented control and eight clinically-diagnosed ADsamples as detected on the FTMS and Q-Star.

B. CSF Biomarkers

The metabolites and their associations with the clinical variablesdescribed in this invention are further confirmed using an independentmass spectrometry system. Representative sample extracts from eachvariable group are re-analyzed by LC-MS using an HP 1050high-performance liquid chromatography, or equivalent interfaced to anABI Q-Star, or equivalent mass spectrometer to obtain mass and intensityinformation for the purpose of identifying metabolites that differ inintensity between the clinical variables under investigation.

Example 3 Structure Elucidation of the Primary Metabolite Biomarkers

Characteristics that can be used for structure elucidation ofmetabolites include accurate mass and molecular formula determination,polarity, acid/base properties, NMR spectra, and MS/MS or MSn spectra.These data, and in particular the MS/MS spectrum, can be used asfingerprints of a particular metabolite and are unique identifiers of aparticular metabolite regardless of whether the complete structure hasbeen determined.

A. Serum Biomarkers—Structural Elucidation

1. LC Retention Time.

The extracts containing the metabolites of interest were subjected toreverse phase LC-MS using a C18 column and analysis by MS as describedin Example 2 above. Table 14 lists the resulting retention times anddetected masses for each of the six serum metabolite markers. Theretention time for all six of the biomarkers is approximately 29-42minutes under these HPLC conditions.

2. Extraction Conditions.

The conditions of extraction also provide insights about the chemicalproperties of the biomarkers. All eight metabolites in the serum (fromExample 1) were ionized in negative mode (3 in APCI and 5 in ESI), whichis indicative of a molecule containing an acidic moiety such as acarboxylic acid or phosphate. Any moiety capable of losing a hydrogenatom can be detected in negative ionization mode. Three of themetabolite markers were extracted into an organic ethyl acetate fraction(plasmalogen metabolites), indicating that these metabolites arenon-polar under acidic condition; one was extracted into an organicethyl acetate fraction dried down and resuspended in butanol, indicatingthat this metabolite (plasmalogen metabolite) is non-polar under acidicconditions. Four of the metabolites (phosphatidyl choline relatedmetabolites) did not extract into the organic fraction, but ratherremained in the aqueous methanol/ammonium hydroxide fraction, indicatingthat these metabolites are very polar.

3. MS/MS Spectra.

The six serum metabolites identified as having the best diagnosticability were subject to MS/MS fragmentation using collision induceddissociation (CID). The structure of a given molecule will dictate aspecific fragmentation pattern under defined conditions that is specificfor that molecule (equivalent to a person's fingerprint). Even slightchanges to the molecule's structure can result in a differentfragmentation pattern. In addition to providing a fingerprint of themolecule's identity, the fragments generated by CID can be used to gaininsights about the structure of a molecule, and for generating a veryspecific high-throughput quantitative detection method (see [30-33] forexamples). FIGS. 6 through 11 show the MS/MS spectra for each of the sixmarkers at −50V collision energy (CE) voltages.

The masses resulting from CID MS/MS of each parent mass were then usedto calculate putative formulas for each of the fragment ions for themetabolites specific to the ADAS-cog scores, as shown in the tables foreach marker (Tables 15 to 17). The information inherent in thefragmentation data is highly specific and descriptive for eachmetabolite, which can be used to gain structural insights about eachmolecule. MS/MS was carried out on the ABI-Q Star XL with all parametersas previously mentioned using Nitrogen as the collision gas at 5 psi andcollision energy (CE) settings of −50 volts.

Based on the fragmentation pattern and masses, the metabolite markersspecific to the ADAS-cog scores have been assigned structures having aphosphatidylcholine-related backbone. From the CID MS/MS, the molecularformulae of 3 metabolites specific to the ADAS-cog scores (accurateneutral masses of 541.3432, 569.3687, 803.568) were determined to beC₂₅H₅₁NO₉P, C₂₇H₅₅NO₉P, and C₄₃H₈₁NO₁₀P, respectively. Their structuresare shown in FIGS. 12-14. The putative structures of additional markersare shown in FIG. 15.

The 3 metabolites specific to AD pathology, with accurate neutral massesof 751.5555, 699.5198, and 723.5195, were analyzed using FT-ICRMS andLC/MS techniques, and by HRAPCI-MS, and MS/MS spectral analysis.Daughter ions determined from the fragmentation pattern for eachmetabolite marker are shown in FIGS. 16-18. The molecular formulae weredetermined to be C₄₃H₇₈NO₇P, C₃₉H₇₄NO₇P, and C₄₁H₇₄NO₇P, respectively.Based on the fragmentation pattern and masses, the metabolite markersspecific to AD pathology have been assigned structures having anethanolamine plasmalogen backbone.

For the 751.5555 metabolite (C₄₃H₇₈NO₇P), and due to negative ionizationconditions, the HRAPCI-MS m/z measured was 750.5482 ([M-H]-, calcd.750.5477 for C₄₃H₇₇NO₇P). The relative intensity of the MS/MS fragmentmasses (MS/MS m/z) were measured as follows: 750 ([M-H]-, 25%), 482(1%), 464 (12%), 446 (5%), 329 (8%), 303 (100%), 259 (12%), 205 (8%),140 (8%). The MS/MS fragments are shown in FIG. 16. The strong MS/MSfragment ion at m/z 303 and other fragment ions due to loss of sn-2 acylgroup (m/z 464) as a ketone, loss of the sn-1 vinyl ether side chain(m/z 482) though small, and the fragment ion due to phosphoethanolamine(m/z 140) indicated the metabolite to be a plasmenylphosphatidylethanolamine-type molecule with arachidonic acid at the sn-2position. Based on these results, the structure of the 751.5555metabolite was elucidated as1-O-1′-(Z)-octadecenyl-2-arachidoyl-sn-glycero-3-phosphoethanolamine.This was confirmed by comparison of their LC/MS and MS/MS spectral data(FIG. 19)

The two remaining metabolites with molecular formulae C₃₉H₇₄NO₇P(neutral mass 699.5198) and C₄₁H₇₄NO₇P (neutral mass 723.5195) werefound to co-elute with the 751.5555 metabolite in LC/MS. Themetabolites' MS/MS fragment ions and fragmentation patterns were similarto those of the 751.5555 metabolite.

For the 699.5198 metabolite, the HRAPCI-MS m/z measured was 698.5125([M-H]-, calcd. 698.5130 for C₃₉H₇₃NO₇P). The relative intensity of theMS/MS m/z were measured as follows: 698 ([M-H]⁻, 8%), 536 (4%), 279(100%), 255 (15%), 119 (10%). The MS/MS fragments are shown in FIG. 17.Based on these results and on its structural similarity to the 751.5555metabolite, the structure of the 699.5198 metabolite was determined tobe 1-O-1′-(Z)-hexadecenyl-2-linoleyl-sn-glycero-3-phosphoethanolamine.

For the 723.5195 metabolite, the HRAPCI-MS m/z measured was 722.5124([M-H]⁻, calcd. 722.5130 for C₄₁H₇₃NO₇P). The relative intensity of theMS/MS m/z were measured as follows: 722 ([M-H]⁻, 12%), 482 (1%), 436(15%), 418 (6%), 303 (100%), 279 (6%), 259 (15%), 255 (10%), 205 (4%),140 (5%). The MS/MS fragments are shown in FIG. 18. Based on theseresults and on its structural similarity to the 751.5555 metabolite, thestructure of the 723.5195 metabolite was proposed as1-0-1′-(Z)-hexadecenyl-2-arachidoyl-sn-glycero-3-phosphoethanolamine.

4. NMR Spectra.

The MS/MS fragmentation provides highly specific descriptive informationabout a metabolite. However, nuclear magnetic resonance (NMR) can assistin solving and confirming the structures of the metabolites. As NMRanalysis techniques are typically less sensitive than mass spectrometrytechniques, multiple injections are performed on the HPLC and theretention time window corresponding to the metabolites of interestcollected and combined. The combined extract is then evaporated todryness and reconstituted in the appropriate solvent for NMR analysis.

Multiple NMR techniques and instruments are available, for example, NMRspectral data are recorded on Bruker Avance 600 MHz spectrometer withcryogenic probe after the chromatographic separation and purification ofthe metabolites of interest. 1H NMR, ¹³C NMR, noe-difference spec, aswell as 2-D NMR techniques like heteronuclear multiple quantumcorrelation (HMQC), and heteronuclear multiple bond correlation (HMBC)are used for structure elucidation work on the biomarkers.

B. CSF Biomarkers

The structural characteristics (LC retention time, extractionconditions, MS/MS fragments) for the 12 CSF metabolite markers aredetermined in the same manner as detailed above.

Example 4 Characterization of Ethanolamine Phospholipids in Serum

Based on the fact that the metabolite markers specific to AD pathologyhave an ethanolamine plasmalogen backbone, it was further investigatedwhether other serum plasmalogens could be indicative of AD. Thischaracterization of ethanolamine phospholipids in serum was made using achromatographic method combined with a mass spectrometric detector.

For MS/MS applications and experiments involving chromatography, anAgilent 1100 HPLC system was used in combination with an AppliedBiosystems QSTAR XL mass spectrometer. An Agilent Zorbax RX-SIL (4.6×150mm, 5 μm) column was used for normal phase chromatography. Conditionsincluded an isocratic mobile phase (55:40:5 isopropanol:hexane:H2O) at aflow rate of 1.0 mL/min for a total run time of 15 min. The column washeated to 35° C. The sample injection volume was 10 μL. Organic solventextracts (ethyl acetate) of samples were evaporated to dryness undernitrogen gas and the residue was reconstituted in 100 μL of 55:40:5isopropanol:hexane:H2O solution prior to injection.

The QSTAR XL instrument was equipped with an APCI (Heated Nebulizer)source operating in negative mode. Values of major instrument parameterswere DP, -60; FP, -265; DP2, -15; GS1, 75; GS2, 15; CUR, 30; NC, -3;TEM, 400° C.; Scan range, 50-1500amu; Accumulation time, I sec.

The three classes of ethanolamine phospholipids are described as diacyl(also referred to herein as PtdEt), alkyl-acyl (also referred to hereinas plasmanyl) or alkenyl-acyl (also referred to herein as EtnPl orplasmenyl). Various basic structures of ethanolamine phospholipids areshown in FIG. 20, along with the standard naming convention used herein.Table 18 shows a list of plasmanyl and plasmenyl ethanolaminephospholipids (M5-M24) that are presently identified and are ofparticular interest.

FIGS. 21-32 show structural information pertaining to selectedmetabolites detected in serum. These figures illustrate the retentiontime, MS/MS fragmentation patterns, and putative structures for selectedmolecules. Due to the conserved MS/MS fragmentation mechanism betweenthese molecules, the theoretical MS/MS transition can be determined forany ethanolamine phospholipid by using a combination of the parent ionmass and the fragment mass of the moiety at either the sn-1 or sn-2position.

Example 5 High Throughput Commercial Method Development

A high throughput method for differential diagnosis of AD dementia andnon-AD dementia states was established.

High throughput screening (HTS) was performed with a linear ion trapmass spectrometer (Q-trap 4000, Applied Biosystem) coupled with Agilent1100 LC system. Sample was prepared by adding 15 uL of internal standard(5 μg/mL of (24-13C)-Cholic Acid in methanol) to 120 uL ethyl acetatefraction of each sample. 100 ul sample was injected by flow injectionanalysis (FIA), and monitored under negative APCI mode. The method wasbased on multiple reaction monitoring (MRM) scan mode of oneparent/daughter transition for each metabolite and one internalstandard. Each transition was scanned for 70 ms for a total cycle timeof 2.475 sec. The isocratic 10% EtOAc in MeOH elution was performed witha flow rate at 360 μl/min for 1 min. The source parameters were set asfollows: CUR: 10.0, CAD: 8, NC: −4.0, TEM: 400, GS1: 30, GS2: 50,interface heater on. The compound parameters were set as follows: DP:−120.0, EP: −10, NC: −4.0, CE: −40, CXP: −15. FIG. 33 illustrates arepresentative standard curve for this method for EtnPls 16:0/22:6generated by diluting a normal serum sample while maintaining a constantconcentration of internal standard (24-13C)-Cholic Acid).

Example 6 Effect of Aging and Severity of Dementia on Serum Levels ofEthanolamine Phospholipids

The effect of aging and severity of dementia on serum levels ofethanolamine phospholipids in 752 subjects aged 40-95 with variouslevels of dementia was investigated. The clinical data on the subjectcohorts is shown in Table 19.

The effect of age was evaluated using a set of aged 30 to 95 subjects ofuntested cognitive status who did not suffer from dementia. Subjectswere divided into one of five subgroups based upon their decade of life(30's, 40's, 50's, 60's, and >70). The 40-49 cohort was used as thepre-dementia reference group due to the low incidence of dementia atthis age. The metabolites of interest (see Table 18) were measured usingthe high throughput method described in Example 5.

The effect of dementia severity was determined in subjects aged 56 to95, comprised of 68 cognitively confirmed non-demented subjects(MMSE≧28); 256 subjects currently diagnosed with SDAT (ADAS-cog 6-70,MMSE 0-26); 20 post-mortem confirmed SDAT and 20 post-mortem confirmedcontrols. Subjects were grouped into one of four dementia subgroupsbased upon either their MMSE score [≧28=Cognitively Normal] or theirADAS-cog score [5-19 =low cognitive impairment); 20-39 =moderate; 40-70=severe].

6A. Absolute Levels of Ethanolamine Phospholipids

A significant gender bias was observed in that only females exhibited anage-related decrease in EtnPls. Free docosohexanoic acid (DHA, Free22:6, M25) in both males and females was significantly increased in the50-59, 60-69, and 70+cohorts relative to the 40-49 cohorts. However,only males exhibited a concomitant increase in both 16:0/22:6-EtnPl(M19) and 18:0/22:6-EtnPl (M24) (see Tables 20-21 for males; Tables22-23 for females). These data indicate that, in females, there may bean age-related dysfunction in the packaging of DHA into EtnPls. Thisgender difference may explain the increased incidence of dementia invery old females (19).

In both males and females, the majority of EtnPls in all dementiasubgroups were significantly reduced relative to cognitive controls. Inboth males and females, free DHA (M25) was significantly decreased onlyin severely demented subjects. In females, a dementia effect wasobserved for three EtnPls (16:0/18:2 (M16), 18:0/18:2 (M21), and16:0/20:4 (M17)) in that both 18:2-containing EtnPls were significantlylower in severely demented subjects versus either low or moderatelydemented females, and 16:0/20:4 (M17) was lower in the severe groupversus the low group (see Tables 24-26). In males, a dementia effect wasobserved for DHA (M25) and 16:0/22:6 (M19) in that free DHA (M25) wasreduced in the moderate group versus the low group and in the severegroup versus the moderate group and 16:0/22:6 (M19) was reduced in thesevere group versus the low group (see Tables 27-29). These resultsindicate that the progressive cognitive deterioration in AD manifestsslightly differently in the two sexes.

Brain white matter contains primarily 18:1- and 18:2-containing EtnPlswith low levels of 20:4-containing and 22:6-containing EtnPls, whereasgray matter contains significantly higher levels of 20:4-containing and22:6-containing EtnPls [34]. In females, increasing dementia appears toaffect both white (18:2) and gray (20:4) matter EtnPls equally, whereasin males predominantly gray (22:6) matter EtnPls appear to be affectedto a greater extent.

Post-mortem collected serum samples from 20 pathologically confirmed ADsubjects and 20 subjects containing minimal amyloid deposition were alsoanalyzed. Both gray and white matter EtnPls were significantly decreasedin post-mortem confirmed AD relative to age matched controls (see Tables30 and 31).

6B. Relative Levels of Ethanolamine Phospholipids

The data collected above was re-analyzed to obtain a ratio between thelevels of each ethanolamine phospholipids with 16:0/18:0 PtdEt (M01).Measurement of the ethanolamine phospholipid levels in this manner ismore stable and reproducible than measurement of the absolute levels.Furthermore, because the 16:0/18:0 PtdEt (M01) is naturally present inall samples and does not appear to vary significantly over diseasestates, this approach minimizes the sample-to-sample variability (due tohandling, extraction, etc).

The results obtained further support the observations and conclusionsmade in 6A. The gender bias was with respect to an age-related decreasein EtnPls was evident in data where ratios to M01 were measured (seeTables 32-33 for males; Tables 34-35 for females). The same trends withrespect to the severity of cognitive impairment were also observed (seeTables 36-38 for males; Tables 39-41 for females). In addition,pathology results on post-mortem serum samples show similar trends(Tables 42 and 43).

Both the absolute EtnPls levels and the EtnPls to M01 ratio exhibited asignificant dementia effect. The EtnPls to M01 ratios of all eightEtnPls (16:0/18:1 (M15), 16:0/18:2 (M16), 16:0/20:4 (M17), 16:0/22:6(M19), 18:0/18:1 (M20), 18:0/18:2 (M21), 18:0/20:4 (M22), 18:0/22:6(M24)) were significantly lower in the severely demented group relativeto the low group while six of the eight were significantly lower in thesevere group relative to the moderate group

Example 7 The Grey and White Matter Score Distribution

A white and gray matter specific EtnPl scoring system was developedwhereby each EtnPl in each subject was normalized to their respectivegender-specific cognitively normal mean, log2 transformed and meancentered. Each subject's white matter score was taken as the lowest suchvalue of plasmenyl 16:0/18:1 (M15), 16:0/18:2 (M16), 18:0/18:1 (M20),and 18:0/18:2 (M21) EtnPls, and their gray matter score as the lowest ofplasmenyl 16:0/20:4 (M17), 16:0/22:6 (M19), 18:0/20:4 (M22), and18:0/22:6 (M24) EtnPls.

These simplified scores revealed that both gray and white matter EtnPlswere decreased at all stages of AD (FIG. 39) and that the levels inpost-mortem confirmed AD closely matched levels in severely dementedsubjects of both sexes (Tables 44-45). The cross-sectional white andgray matter score distributions in subjects of various levels ofdementia clearly showed a dementia dependent shift in the populationmeans (Tables 46-47). The effect of age on the white and gray matterscores was also determined (Tables 48-49). This also indicated thatchanges in serum levels of gray matter EtnPls may precede white matterchanges and potentially be an early risk factor for AD. Suchcross-sectional data does not account for baseline variability amongsubjects. Individual longitudinal trajectories of these scores may bemore accurate at detecting early risk of AD in otherwise healthy,non-demented subjects.

Based on these scores, risk prediction can be performed on both male andfemale subjects (Tables 49-50) where a cut-off value that results inapproximately 20-30% of cognitively normal subjects being classified aseither intermediate or high risk is used. Using this cut-off value, asubject's white and gray matter score is evaluated. If the subject testsnormal on both scores, the subject is deemed to be at low risk. If thesubject tests positive on one of the scores, the subjects is deemed tobe at intermediate risk and if the subject tests positive on bothscores, the subject is deemed to be at high risk.

Example 8 Effect of Dementia Severity and AD Pathology on Serum EtnPlsLevels in Combined Male and Female Subjects

The effect of dementia severity was determined using 324 subjects (176female, 148 male) aged 56 to 95, comprised of 68 cognitively confirmednon-demented subjects (MMSE≧28) and 256 subjects currently diagnosedwith AD (ADAS-cog 6-70, MMSE 0-26). The effect of AD pathology wasdetermined using serum samples collected from 20 post-mortem confirmedAD and 19 control subjects (Table 19). Subjects were grouped into one offour dementia severity cohorts based upon either their MMSE score [≧28=Cognitively Normal] or their ADAS-cog score [5-19 =low cognitiveimpairment; 20-39 =moderate; 40-70 =severe].

Mean serum levels of 16:0/18:1 (M15), 16:0/18:2 (M16), 16:0/20:4 (M17),16:0/22:6 (M19), 18:0/18:1 (M20), 18:0/18:2 (M21), 18:0/20:4 (M22),18:0/22:6 (M24) EtnPls; free docosahexaenoic acid (DHA, Free 22:6, M25);and phosphatidylethanolamine (PtdEt) 16:0/18:0 (D16:0/18:0; M01) weredetermined for each group (FIG. 34). All eight EtnPls in all dementiasubgroups were observed to be significantly reduced relative tocognitive controls (24 pair-wise comparisons, t-test p-values 2.6e-2 to2.0e-10, median =3.9e-5). Free DHA (M25) was significantly decreased inboth moderately and severely demented subjects (p<0.05). All eightEtnPls were also significantly decreased in post-mortem confirmed SDATrelative to age matched controls. D16:0/18:0 (M01) levels, anon-plasmalogen phoshopholipid remained unchanged across the differentdementia cohorts.

Example 9 Population Distributions as a Function of Dementia Severity

The EtnPls 16:0/22:6 (M19) to PtdEt 16:0/18:0 ratio (M01) (DHA-EtnPls)showed the strongest overall sex-independent dementia effect (Tables 38,41) and was used for all subsequent population distributions andcomparisons. A summary of the key comparisons using this ratio arelisted in Table 52. This ratio was then log(2) transformed and used tocreate a population histogram for each cohort of increasing cognitiveimpairment (FIG. 35). A cut-off value was selected based upon thefindings of Bennett et al [35], (i.e. ˜30% of the CN group beingdetected as AD) (FIG. 35, dotted line). Using this cut-off, 63%, 79% and83% of low, moderate and severely demented subjects, respectively, weresubsequently classified as AD.

To compare these distributions with the known distributions of Aβpathology in AD, the results of four prospective pathology studies [8,35-37] were combined to generate the theoretical populationdistributions of Aβ pathology in demented and non-demented populations,assuming that Aβ is normally distributed in each population (FIG. 36A).These studies reported that only 71% (140/198) of clinically diagnosedAD subjects have AD pathology at death and that 32% (87/268) ofcognitively normal subjects meet neuropathological criteria for AD atdeath. When the data from all of our cognitively tested subjects werecombined, 32% (22/68) of our non-demented population and 75% (192/256)of our demented population were classified as AD positive based upontheir serum EtnPls level (FIG. 36B). This comparison revealed that theobserved distribution of depleted 22:6-containing EtnPls perfectlymatched the theoretical distribution of AD pathology in demented andnon-demented subjects.

Example 10 Linear Extrapolation of Disease Progression and Serum EtnPlsDepletion

To investigate whether a correlation between the decrease in EtnPls andincreasing dementia in the clinically diagnosed AD population exists, alinear regression analysis was performed using the mean 22:6-containingEtnPls level (normalized to CN) of each of the dementia cohorts and theaverage ADAS-cog score for each of these three cohorts (FIG. 37). A veryhigh correlation was observed between the mean 22:6-containing EtnPlslevel and the mean ADAS-cog scores of the three dementia cohorts(r2=0.99). However, this linear decrease did not extrapolate back to theCN group (X vs. CN). Assuming a clinical AD progression of 7.5 ADAS-cogunits per year, this extrapolation predicts that that 22:6-containingEtnPls levels begin to decline approximately seven years before clinicalcognitive impairment (ADAS-cog =15) is evident.

Example 11 The Effect of Chronological Age on Serum DHA-EtnPls Levels

To investigate whether the above prediction could be verifiedexperimentally, the serum 22:6-containing EtnPls levels in 209 subjects(110 male, 99 female, Table 19) of unknown cognitive status butcurrently not diagnosed with dementia was determined and compared to theclinical AD and CN cohorts (FIG. 38). The results of this analysisrevealed a significant drop in serum 22:6-containing EtnPls in the aged60-69 cohort versus the aged 50-59 cohort (FIG. 38A). This cohort alsohad significantly lower levels versus the CN group even though the CNgroup was, on average, 13 years older. Interestingly, the aged 70-95cohort was not significantly different from either the aged 50-59 cohortor the CN cohort, but had significantly higher levels than the SDATcohort.

Example 12 Sub-Populations Identified by Serum DHA-EtnPls Levels

The distribution of serum DHA-EtnPls within each age group, as shown inFIG. 38B, was also examined. The population distributions of the fivegroups (three age groups, CN and AD) differentiated by age and dementiastatus reveal the presence of three distinct populations (P1-P3, FIG.38B). The populations were assigned as: P1—subjects with AD pathologyand no remaining reserve capacity; P3—subjects with little or no ADpathology; P2—subjects that are transitioning from P3 to P1. These P2subjects are hypothesized to have AD pathology and some level of reserveremaining.

Since AD subjects have a life expectancy of less than 10 years fromdiagnosis [38, 39] and low 22:6-containing EtnPls are highly associatedwith AD severity, the decreased number of P1 subjects observed in theaged 70-95 cohort is most likely due to differences in life expectancybetween P1 and P2 or P3. The transitory nature of P2 is best illustratedby examining the different ratios between the percentages of subjectspresent in P3 compared to P2, as observed in the lower three panels ofFIG. 7B. These three cohorts differ only in dementia status. The P3 toP2 ratio changes from 3:1 (68% versus 22%) in the confirmed cognitivenormal group to an intermediate ratio of 1:1 (43% versus 46%) in thenormal healthy elderly group of unknown cognitive status, to 0.6:1 (25%versus 40%) in the confirmed demented AD cohort.

All citations are hereby incorporated by reference.

The present invention has been described with regard to one or moreembodiments. However, it will be apparent to persons skilled in the artthat a number of variations and modifications can be made withoutdeparting from the scope of the invention as defined in the claims.

TABLE 1 Accurate mass features differing between clinically diagnosed ADpatients and non-demented controls (p < 0.05, log2 transformed).Detected Analysis AVG (log2) SEM AVG (log2) SEM log(2) Mass Mode AD ADNormal Normal Ratio P Value 723.5197 1204 3.576 0.039 4.350 0.056 0.8225.09E−19 723.5195 1202 2.186 0.040 2.892 0.052 0.756 4.86E−17 724.52571204 2.866 0.032 3.463 0.055 0.828 4.15E−16 749.5367 1202 3.176 0.0343.714 0.041 0.855 3.82E−15 751.5555 1204 4.575 0.041 5.248 0.060 0.8721.07E−14 751.5529 1202 3.335 0.036 3.920 0.050 0.851 1.14E−14 752.55641202 2.251 0.038 2.836 0.050 0.794 3.13E−14 752.5583 1204 3.472 0.0424.094 0.061 0.848 6.47E−13 699.5198 1204 2.216 0.038 2.775 0.054 0.7996.76E−13 750.544 1204 3.279 0.038 3.858 0.063 0.850 1.07E−12 749.54071204 4.426 0.039 5.012 0.064 0.883 1.33E−12 541.3432 1102 3.315 0.0333.798 0.048 0.873 1.42E−12 750.5402 1202 2.192 0.038 2.704 0.044 0.8112.76E−12 725.5385 1204 2.884 0.043 3.417 0.054 0.844 1.28E−10 569.36871102 2.262 0.039 2.724 0.048 0.830 6.22E−10 727.5568 1204 3.518 0.0383.986 0.060 0.882 1.70E−09 804.5713 1102 4.207 0.042 4.610 0.033 0.9133.88E−08 803.568 1102 5.432 0.043 5.838 0.035 0.930 7.10E−08 726.54611204 2.808 0.032 3.150 0.050 0.892 7.50E−08 827.57 1102 4.151 0.0484.630 0.062 0.897 9.42E−08 803.5445 1101 5.123 0.055 5.655 0.059 0.9061.01E−07 555.3102 1102 1.818 0.045 2.240 0.046 0.812 1.27E−07 565.33941102 3.480 0.050 3.958 0.055 0.879 1.28E−07 804.5476 1101 4.169 0.0564.703 0.060 0.887 1.33E−07 828.5737 1102 3.138 0.046 3.590 0.061 0.8741.89E−07 567.3547 1102 2.822 0.041 3.218 0.054 0.877 2.89E−07 728.56271204 2.935 0.033 3.281 0.060 0.895 5.16E−07 817.5377 1102 2.282 0.0482.712 0.057 0.842 8.85E−07 779.5444 1101 6.433 0.053 6.874 0.043 0.9361.06E−06 780.5474 1101 5.437 0.053 5.875 0.043 0.925 1.28E−06 812.57621202 1.659 0.050 2.084 0.058 0.796 2.24E−06 832.6026 1102 3.455 0.0413.795 0.040 0.910 2.48E−06 811.5732 1202 2.705 0.036 3.027 0.055 0.8933.40E−06 871.5528 1102 3.068 0.042 3.408 0.040 0.900 3.47E−06 831.59971102 4.564 0.042 4.903 0.040 0.931 3.48E−06 793.5386 1102 3.604 0.0433.950 0.039 0.912 3.50E−06 782.5085 1204 3.401 0.045 3.780 0.055 0.9003.81E−06 805.5832 1102 4.075 0.047 4.485 0.068 0.909 3.87E−06 781.56171101 6.109 0.061 6.610 0.072 0.924 5.14E−06 813.5885 1202 3.012 0.0303.276 0.048 0.919 6.23E−06 794.5421 1102 2.523 0.042 2.853 0.040 0.8856.30E−06 814.5917 1202 2.041 0.026 2.289 0.051 0.892 7.46E−06 747.52451204 3.473 0.043 3.886 0.090 0.894 9.55E−06 837.5027 1101 3.578 0.0453.933 0.050 0.910 1.02E−05 782.565 1101 5.083 0.063 5.589 0.078 0.9091.09E−05 746.5717 1204 3.085 0.031 3.362 0.061 0.918 1.82E−05 829.58561102 4.043 0.048 4.398 0.046 0.919 1.85E−05 784.5237 1204 3.310 0.0403.603 0.037 0.919 1.91E−05 786.5416 1204 3.815 0.035 4.087 0.043 0.9331.91E−05 760.5216 1204 4.075 0.036 4.347 0.039 0.938 2.11E−05 745.56581204 3.937 0.034 4.242 0.068 0.928 2.12E−05 744.5536 1204 4.322 0.0344.605 0.058 0.939 2.46E−05 783.5672 1101 3.755 0.068 4.259 0.079 0.8823.48E−05 807.5758 1101 5.736 0.052 6.102 0.047 0.940 3.69E−05 808.57921101 4.697 0.052 5.063 0.047 0.928 4.20E−05 743.5471 1204 5.286 0.0365.579 0.063 0.947 4.94E−05 482.3215 1202 1.971 0.038 2.251 0.062 0.8750.0001 755.486 1204 3.221 0.047 3.561 0.057 0.905 0.0001 758.5092 12044.574 0.033 4.808 0.042 0.951 0.0001 775.5533 1202 2.120 0.044 2.4490.068 0.866 0.0001 787.5729 1202 1.847 0.040 2.145 0.057 0.861 0.0001795.5181 1101 2.630 0.059 3.044 0.064 0.864 0.0001 795.555 1102 2.6650.043 2.986 0.065 0.892 0.0001 805.5605 1101 5.414 0.057 5.785 0.0480.936 0.0001 831.5759 1101 4.297 0.056 4.677 0.060 0.919 0.0001 855.60161102 3.538 0.045 3.873 0.065 0.914 0.0001 517.314 1101 5.470 0.038 5.7550.069 0.951 0.0002 541.3139 1101 4.091 0.053 4.494 0.096 0.910 0.0002542.3173 1101 2.284 0.055 2.687 0.089 0.850 0.0002 747.5201 1202 1.9370.051 2.313 0.088 0.838 0.0002 757.4991 1101 3.644 0.065 4.073 0.0700.895 0.0002 775.5528 1204 3.197 0.045 3.537 0.085 0.904 0.0002 806.56391101 4.423 0.057 4.779 0.049 0.926 0.0002 832.5791 1101 3.357 0.0553.723 0.061 0.902 0.0002 915.5191 1101 2.376 0.051 2.717 0.063 0.8740.0002 755.5468 1101 2.326 0.064 2.753 0.085 0.845 0.0003 777.553 12021.859 0.072 2.384 0.130 0.780 0.0003 829.5604 1101 3.535 0.053 3.8750.058 0.912 0.0003 518.3174 1101 3.438 0.038 3.717 0.073 0.925 0.0004731.5464 1101 1.945 0.096 2.600 0.157 0.748 0.0004 757.5626 1101 6.6550.074 7.108 0.079 0.936 0.0004 758.5656 1101 5.702 0.075 6.160 0.0780.926 0.0004 759.5779 1101 5.547 0.069 5.997 0.099 0.925 0.0004 760.58111101 4.419 0.071 4.886 0.103 0.904 0.0004 768.5539 1204 3.964 0.0464.279 0.077 0.927 0.0004 748.5287 1204 2.277 0.091 2.820 0.088 0.8070.0005 783.5148 1204 3.284 0.047 3.574 0.056 0.919 0.0005 821.5712 11023.064 0.040 3.304 0.044 0.927 0.0005 523.4679 1203 3.252 0.121 4.0110.165 0.811 0.0006 781.562 1201 7.364 0.040 7.638 0.071 0.964 0.0006810.5399 1204 2.835 0.050 3.145 0.066 0.901 0.0006 732.4938 1204 4.2490.042 4.511 0.057 0.942 0.0007 522.4635 1203 4.713 0.126 5.485 0.1760.859 0.0008 810.5969 1101 3.957 0.072 4.370 0.080 0.905 0.0009 853.58541102 2.626 0.043 2.880 0.052 0.912 0.0009 819.5551 1102 2.394 0.0462.654 0.046 0.902 0.001 828.5743 1202 5.243 0.058 5.598 0.085 0.9360.001 478.254 1201 1.584 0.083 1.051 0.148 1.507 0.0011 579.5325 12034.102 0.089 4.636 0.127 0.885 0.0011 744.4956 1204 3.881 0.039 4.1170.058 0.943 0.0011 761.5843 1101 2.422 0.065 2.829 0.106 0.856 0.0011809.5936 1101 4.964 0.070 5.362 0.079 0.926 0.0011 886.5582 1102 3.0410.037 3.250 0.041 0.936 0.0011 481.3172 1202 3.923 0.039 4.159 0.0600.943 0.0012 767.5495 1204 5.063 0.049 5.369 0.082 0.943 0.0012 782.56531201 6.339 0.043 6.614 0.076 0.959 0.0012 827.5701 1202 6.306 0.0636.684 0.092 0.943 0.0012 847.5316 1101 2.742 0.059 3.091 0.083 0.8870.0012 789.5892 1202 1.723 0.036 1.952 0.065 0.883 0.0013 543.3296 11013.733 0.044 4.023 0.087 0.928 0.0014 575.2728 1101 2.470 0.043 2.7390.074 0.902 0.0014 580.5351 1203 1.456 0.127 2.192 0.174 0.664 0.0014521.4522 1203 2.302 0.131 3.028 0.158 0.760 0.0016 731.4916 1204 5.4820.046 5.746 0.063 0.954 0.0016 759.5163 1204 4.724 0.042 4.954 0.0480.954 0.0016 306.2569 1204 2.998 0.046 3.266 0.067 0.918 0.0017 771.58141204 4.164 0.036 4.367 0.046 0.954 0.0017 786.5967 1101 4.808 0.0745.203 0.078 0.924 0.0019 458.2405 1101 1.736 0.040 1.986 0.077 0.8740.0021 520.4499 1203 3.956 0.112 4.577 0.148 0.864 0.0021 748.5735 12023.918 0.035 3.722 0.050 1.053 0.0021 490.3641 1203 1.944 0.092 1.3970.159 1.391 0.0023 545.3453 1101 3.606 0.051 3.898 0.078 0.925 0.0023605.5457 1203 5.135 0.068 5.509 0.089 0.932 0.0023 769.5656 1204 3.9630.038 4.174 0.054 0.950 0.0023 570.3725 1202 2.976 0.032 3.155 0.0450.943 0.0024 785.5933 1101 5.884 0.074 6.271 0.080 0.938 0.0024 582.24731201 3.325 0.096 2.793 0.141 1.191 0.0026 569.369 1202 4.908 0.033 5.0890.047 0.964 0.0027 784.5811 1101 4.405 0.079 4.817 0.089 0.915 0.0027811.6096 1101 3.078 0.083 3.511 0.096 0.877 0.0027 590.343 1202 4.0250.050 4.304 0.076 0.935 0.0028 856.672 1202 2.764 0.038 2.553 0.0571.082 0.0028 833.5932 1101 3.276 0.068 3.608 0.056 0.908 0.003 506.28511201 3.142 0.077 2.656 0.165 1.183 0.0031 793.5681 1204 3.155 0.0403.372 0.055 0.936 0.0031 546.3485 1101 1.999 0.050 2.287 0.089 0.8740.0036 591.3542 1202 4.045 0.061 4.345 0.058 0.931 0.0037 741.5305 12042.931 0.056 3.250 0.102 0.902 0.0042 796.5876 1204 2.634 0.043 2.8600.062 0.921 0.0042 804.7227 1203 1.842 0.145 2.611 0.220 0.705 0.0044807.59 1202 2.463 0.045 2.718 0.082 0.906 0.0045 506.3213 1202 2.5380.040 2.748 0.061 0.923 0.0046 552.5022 1203 3.164 0.088 3.643 0.1470.869 0.0047 589.3403 1202 5.876 0.056 6.171 0.085 0.952 0.0048 806.58731202 4.366 0.047 4.635 0.092 0.942 0.0048 550.4957 1203 6.898 0.0967.415 0.160 0.930 0.0049 604.5433 1203 6.554 0.069 6.901 0.089 0.9500.005 805.5839 1202 5.562 0.048 5.841 0.097 0.952 0.0052 551.4986 12035.480 0.095 5.988 0.158 0.915 0.0053 743.5469 1202 3.061 0.055 3.3480.086 0.914 0.0056 541.3435 1202 5.669 0.059 5.974 0.094 0.949 0.0058183.0661 1101 2.590 0.091 3.015 0.094 0.859 0.006 858.6212 1202 2.7070.059 2.994 0.074 0.904 0.0061 614.4914 1203 2.779 0.060 2.448 0.1141.135 0.0062 787.5465 1204 2.675 0.077 3.017 0.054 0.887 0.0062 772.58621204 3.287 0.031 3.437 0.042 0.956 0.007 837.5881 1202 2.429 0.027 2.5770.053 0.942 0.007 509.3493 1202 2.403 0.035 2.579 0.055 0.931 0.0071529.3167 1202 3.032 0.048 3.265 0.069 0.928 0.0075 564.5134 1203 2.7060.075 3.075 0.111 0.880 0.0075 566.3434 1202 5.203 0.049 5.436 0.0620.957 0.0075 833.7571 1203 2.962 0.109 3.507 0.170 0.845 0.0077 631.6281203 1.795 0.127 2.391 0.161 0.751 0.008 857.6186 1202 3.773 0.058 4.0490.076 0.932 0.008 858.6861 1202 2.943 0.040 2.756 0.052 1.068 0.0081519.3321 1101 3.964 0.084 4.382 0.133 0.905 0.0083 685.26 1202 1.7710.048 1.998 0.063 0.886 0.0083 757.5014 1204 3.755 0.048 3.971 0.0520.946 0.0085 744.55 1202 1.968 0.051 2.222 0.083 0.886 0.0086 671.57231204 2.231 0.084 2.604 0.087 0.857 0.0087 304.241 1204 4.887 0.041 5.0880.066 0.961 0.0092 536.4794 1203 2.320 0.101 2.799 0.143 0.829 0.0093542.3461 1202 3.873 0.049 4.106 0.074 0.943 0.0095 675.6377 1204 3.9530.043 4.160 0.066 0.950 0.0098 520.3354 1101 2.240 0.085 2.646 0.1300.846 0.01 832.7523 1203 3.859 0.107 4.374 0.169 0.882 0.0103 409.02081202 2.806 0.035 2.980 0.063 0.942 0.0106 768.5503 1202 1.942 0.0762.277 0.090 0.853 0.0111 303.1079 1202 5.648 0.034 5.802 0.047 0.9730.0113 592.3571 1202 2.291 0.063 2.560 0.062 0.895 0.0115 837.718 12042.888 0.176 3.662 0.217 0.788 0.0121 832.7492 1204 3.286 0.119 3.8480.190 0.854 0.0125 832.6037 1202 5.066 0.047 5.274 0.062 0.961 0.013411.3212 1202 2.868 0.038 3.033 0.048 0.946 0.0134 838.7226 1204 2.3130.148 2.973 0.206 0.778 0.0136 670.569 1204 3.239 0.061 3.515 0.0930.921 0.0141 795.5838 1204 3.566 0.046 3.769 0.063 0.946 0.0141 767.5471202 3.073 0.064 3.358 0.092 0.915 0.0143 305.2438 1204 2.519 0.0442.719 0.067 0.926 0.0146 505.3229 1202 3.994 0.051 4.222 0.076 0.9460.0156 803.5677 1202 7.196 0.070 7.502 0.098 0.959 0.0157 711.2577 12022.250 0.049 2.454 0.056 0.917 0.0159 827.5448 1101 3.549 0.077 3.8730.095 0.916 0.016 548.4815 1203 7.094 0.072 7.405 0.103 0.958 0.0174568.3573 1202 4.008 0.035 4.167 0.060 0.962 0.0175 578.5277 1203 4.3010.303 5.548 0.355 0.775 0.0175 601.5164 1203 7.640 0.038 7.463 0.0691.024 0.0185 549.4845 1203 5.666 0.077 5.994 0.110 0.945 0.0187 743.54661203 1.987 0.064 2.266 0.099 0.877 0.0189 772.5278 1204 3.324 0.0393.488 0.055 0.953 0.019 765.5334 1204 3.269 0.060 3.540 0.104 0.9230.0193 440.3532 1204 1.417 0.096 0.975 0.180 1.453 0.0205 495.332 11015.251 0.073 5.565 0.112 0.944 0.0205 804.5718 1202 5.877 0.057 6.1170.080 0.961 0.0206 340.2976 1203 1.597 0.082 1.937 0.112 0.825 0.0208856.6061 1202 4.565 0.056 4.805 0.086 0.950 0.0212 584.2646 1204 3.1360.115 2.649 0.172 1.184 0.0218 733.6426 1204 2.978 0.049 2.718 0.1261.096 0.0219 588.4731 1203 2.387 0.064 2.031 0.179 1.175 0.0223 765.53131202 1.802 0.073 2.098 0.095 0.859 0.0226 523.3634 1101 3.466 0.0823.791 0.103 0.914 0.0235 830.5894 1202 4.847 0.051 5.051 0.064 0.9600.0236 887.7352 1204 6.295 0.082 5.963 0.113 1.056 0.0244 598.5124 12042.094 0.134 1.513 0.233 1.385 0.0249 616.5052 1203 4.205 0.062 3.9570.084 1.062 0.025 916.7743 1204 5.894 0.081 5.571 0.107 1.058 0.025430.3818 1204 4.938 0.075 5.253 0.122 0.940 0.0262 855.6023 1202 5.5520.056 5.784 0.087 0.960 0.0266 684.5489 1204 2.523 0.055 2.745 0.0800.919 0.0269 831.6001 1202 6.192 0.053 6.404 0.074 0.967 0.0269 826.70691204 2.482 0.089 2.819 0.099 0.880 0.0275 915.7681 1204 6.304 0.0855.972 0.116 1.056 0.0293 615.3539 1202 2.463 0.043 2.629 0.058 0.9370.0296 431.386 1204 3.491 0.075 3.795 0.118 0.920 0.0298 942.7879 12043.487 0.078 3.195 0.092 1.091 0.0302 665.501 1204 3.388 0.071 3.0740.142 1.102 0.0305 861.7806 1203 2.531 0.099 2.902 0.119 0.872 0.0306618.4829 1201 1.791 0.120 1.310 0.185 1.367 0.0309 830.7352 1204 4.2520.096 4.638 0.150 0.917 0.031 801.555 1202 2.660 0.056 2.881 0.082 0.9230.0311 739.5143 1204 2.754 0.087 3.121 0.158 0.883 0.0317 492.3816 12033.513 0.055 3.302 0.077 1.064 0.0339 741.5319 1202 1.106 0.080 1.4240.125 0.777 0.0339 914.7583 1204 5.082 0.083 4.763 0.118 1.067 0.0339507.3316 1202 2.940 0.035 3.094 0.071 0.950 0.034 504.3814 1203 1.7010.070 1.413 0.124 1.204 0.0341 496.3355 1101 3.442 0.074 3.736 0.1190.921 0.0347 521.3477 1101 3.717 0.075 4.021 0.127 0.925 0.0351 829.58591202 5.976 0.056 6.181 0.070 0.967 0.0353 686.4877 1204 2.700 0.0512.881 0.052 0.937 0.0358 888.7394 1204 5.702 0.080 5.400 0.110 1.0560.0358 825.6926 1203 1.840 0.110 2.235 0.124 0.823 0.0369 746.557 12022.166 0.030 2.057 0.037 1.053 0.0378 757.5625 1201 7.728 0.046 7.9090.077 0.977 0.0391 615.4798 1204 2.647 0.049 2.827 0.067 0.936 0.0396831.7408 1203 4.104 0.085 4.425 0.130 0.928 0.0405 761.5846 1201 3.1550.051 3.357 0.091 0.940 0.0414 581.3344 1202 1.927 0.076 2.207 0.1100.873 0.0423 1098.9739 1204 2.955 0.090 2.606 0.152 1.134 0.0427380.3096 1204 1.650 0.070 1.899 0.090 0.869 0.0434 565.3394 1202 7.0010.052 7.186 0.068 0.974 0.0439 478.3664 1203 1.493 0.068 1.212 0.1431.232 0.0475 835.7006 1204 2.799 0.115 3.214 0.167 0.871 0.0485 320.23561204 1.447 0.070 1.686 0.085 0.858 0.0486 493.385 1203 1.979 0.066 1.7220.124 1.149 0.049 512.4082 1204 2.415 0.121 1.954 0.215 1.236 0.0493610.3686 1201 5.156 0.083 4.771 0.223 1.081 0.0495 760.5811 1201 5.2250.052 5.422 0.091 0.964 0.0495 600.5127 1203 8.858 0.044 8.692 0.0741.019 0.0496 715.5167 1204 2.592 0.082 2.902 0.141 0.893 0.0498 759.57791201 6.347 0.051 6.541 0.090 0.970 0.0499

TABLE 2 Accurate mass features differing between clinically diagnosed ADpatients with a significant cognitive impairment and non-dementedcontrols (p < 0.05, log2 transformed). Detected Analysis AVG (log2) SEMAVG (log2) SEM log(2) Mass Mode AD + Cog AD + Cog Normal Normal Ratio PValue 541.3432 1102 3.138 0.034 3.798 0.048 1.210 3.45E−17 569.3687 11022.063 0.042 2.724 0.048 1.320 1.67E−15 723.5197 1204 3.581 0.053 4.3500.056 1.215 1.04E−14 803.568 1102 5.238 0.048 5.838 0.035 1.115 2.00E−14804.5713 1102 4.017 0.048 4.610 0.033 1.148 2.02E−14 723.5195 1202 2.1930.056 2.892 0.052 1.319 4.37E−13 749.5367 1202 3.187 0.044 3.714 0.0411.165 2.12E−12 555.3102 1102 1.634 0.053 2.240 0.046 1.371 3.10E−12565.3394 1102 3.265 0.058 3.958 0.055 1.212 3.23E−12 724.5257 1204 2.8500.048 3.463 0.055 1.215 4.54E−12 699.5198 1204 2.129 0.055 2.775 0.0541.303 9.18E−12 871.5528 1102 2.885 0.049 3.408 0.040 1.181 2.08E−11567.3547 1102 2.629 0.050 3.218 0.054 1.224 2.92E−11 751.5555 1204 4.5910.056 5.248 0.060 1.143 3.18E−11 780.5474 1101 5.241 0.064 5.875 0.0431.121 3.70E−11 752.5564 1202 2.281 0.049 2.836 0.050 1.244 4.78E−11779.5444 1101 6.239 0.065 6.874 0.043 1.102 5.01E−11 829.5856 1102 3.8060.059 4.398 0.046 1.156 9.29E−11 794.5421 1102 2.364 0.048 2.853 0.0401.206 1.01E−10 793.5386 1102 3.435 0.052 3.950 0.039 1.150 1.05E−10831.5997 1102 4.372 0.055 4.903 0.040 1.121 1.61E−10 832.6026 1102 3.2680.055 3.795 0.040 1.161 1.67E−10 751.5529 1202 3.369 0.052 3.920 0.0501.163 1.89E−10 749.5407 1204 4.423 0.047 5.012 0.064 1.133 1.93E−10827.57 1102 3.984 0.059 4.630 0.062 1.162 1.95E−10 725.5385 1204 2.8150.058 3.417 0.054 1.214 2.58E−10 804.5476 1101 3.987 0.073 4.703 0.0601.180 2.67E−10 837.5027 1101 3.402 0.050 3.933 0.050 1.156 2.76E−10803.5445 1101 4.941 0.074 5.655 0.059 1.144 3.46E−10 752.5583 1204 3.4840.056 4.094 0.061 1.175 3.59E−10 828.5737 1102 2.982 0.056 3.590 0.0611.204 3.85E−10 750.544 1204 3.258 0.054 3.858 0.063 1.184 6.22E−10750.5402 1202 2.210 0.052 2.704 0.044 1.224 1.19E−09 805.5832 1102 3.8740.056 4.485 0.068 1.158 1.95E−09 727.5568 1204 3.445 0.055 3.986 0.0601.157 7.95E−09 807.5758 1101 5.531 0.069 6.102 0.047 1.103 8.66E−09805.5605 1101 5.184 0.074 5.785 0.048 1.116 9.26E−09 808.5792 1101 4.4880.071 5.063 0.047 1.128 1.11E−08 806.5639 1101 4.199 0.075 4.779 0.0491.138 3.65E−08 915.5191 1101 2.189 0.058 2.717 0.063 1.241 5.21E−08817.5377 1102 2.175 0.064 2.712 0.057 1.247 5.29E−08 781.5617 1101 5.9280.084 6.610 0.072 1.115 8.47E−08 819.5551 1102 2.198 0.058 2.654 0.0461.208 9.28E−08 726.5461 1204 2.759 0.044 3.150 0.050 1.141 1.31E−07783.5672 1101 3.535 0.092 4.259 0.079 1.205 1.69E−07 886.5582 1102 2.9110.041 3.250 0.041 1.117 2.03E−07 782.565 1101 4.901 0.088 5.589 0.0781.140 2.35E−07 784.5237 1204 3.211 0.056 3.603 0.037 1.122 4.12E−07853.5854 1102 2.449 0.056 2.880 0.052 1.176 4.56E−07 795.555 1102 2.5200.054 2.986 0.065 1.185 5.49E−07 855.6016 1102 3.386 0.060 3.873 0.0651.144 6.32E−07 821.5712 1102 2.919 0.053 3.304 0.044 1.132 6.68E−07795.5181 1101 2.458 0.082 3.044 0.064 1.238 7.57E−07 832.5791 1101 3.1840.076 3.723 0.061 1.169 9.68E−07 786.5416 1204 3.729 0.049 4.087 0.0431.096 1.02E−06 831.5759 1101 4.123 0.080 4.677 0.060 1.134 1.03E−06728.5627 1204 2.875 0.047 3.281 0.060 1.141 1.07E−06 757.4991 1101 3.4500.089 4.073 0.070 1.181 1.08E−06 760.5216 1204 4.008 0.048 4.347 0.0391.084 1.13E−06 829.5604 1101 3.360 0.075 3.875 0.058 1.153 1.41E−06847.5316 1101 2.564 0.060 3.091 0.083 1.205 1.84E−06 755.5468 1101 2.1150.086 2.753 0.085 1.302 1.88E−06 782.5085 1204 3.369 0.056 3.780 0.0551.122 2.13E−06 755.486 1204 3.146 0.061 3.561 0.057 1.132 5.93E−06833.5932 1101 3.045 0.095 3.608 0.056 1.185 7.05E−06 758.5656 1101 5.5110.107 6.160 0.078 1.118 1.11E−05 758.5092 1204 4.504 0.047 4.808 0.0421.068 1.24E−05 757.5626 1101 6.470 0.106 7.108 0.079 1.099 1.48E−05760.5811 1101 4.216 0.099 4.886 0.103 1.159 1.48E−05 744.5536 1204 4.2600.047 4.605 0.058 1.081 1.55E−05 759.5779 1101 5.352 0.096 5.997 0.0991.121 1.60E−05 811.6096 1101 2.801 0.116 3.511 0.096 1.253 1.84E−05731.5464 1101 1.634 0.139 2.600 0.157 1.591 1.89E−05 812.5762 1202 1.7520.045 2.084 0.058 1.190 1.95E−05 743.5471 1204 5.227 0.047 5.579 0.0631.067 2.40E−05 784.5811 1101 4.161 0.112 4.817 0.089 1.158 3.03E−05632.5762 1203 0.925 0.149 1.830 0.132 1.979 3.07E−05 761.5843 1101 2.2410.084 2.829 0.106 1.262 4.28E−05 517.314 1101 5.399 0.050 5.755 0.0691.066 0.0001 518.3174 1101 3.358 0.051 3.717 0.073 1.107 0.0001 591.35421202 3.870 0.092 4.345 0.058 1.123 0.0001 732.4938 1204 4.186 0.0564.511 0.057 1.078 0.0001 745.5658 1204 3.881 0.052 4.242 0.068 1.0930.0001 746.5717 1204 3.047 0.043 3.362 0.061 1.103 0.0001 747.5245 12043.433 0.058 3.886 0.090 1.132 0.0001 777.553 1202 1.720 0.100 2.3840.130 1.386 0.0001 783.5148 1204 3.217 0.063 3.574 0.056 1.111 0.0001785.5933 1101 5.687 0.112 6.271 0.080 1.103 0.0001 786.5967 1101 4.6110.112 5.203 0.078 1.128 0.0001 809.5936 1101 4.786 0.104 5.362 0.0791.120 0.0001 810.5969 1101 3.779 0.108 4.370 0.080 1.157 0.0001 811.57321202 2.729 0.047 3.027 0.055 1.109 0.0001 858.6212 1202 2.533 0.0802.994 0.074 1.182 0.0001 306.2569 1204 2.878 0.070 3.266 0.067 1.1350.0002 482.3215 1202 1.940 0.050 2.251 0.062 1.160 0.0002 522.4635 12034.489 0.181 5.485 0.176 1.222 0.0002 523.4679 1203 3.035 0.180 4.0110.165 1.322 0.0002 579.5325 1203 3.962 0.112 4.636 0.127 1.170 0.0002759.5163 1204 4.645 0.061 4.954 0.048 1.066 0.0002 787.5729 1202 1.8340.054 2.145 0.057 1.170 0.0002 810.5399 1204 2.760 0.071 3.145 0.0661.140 0.0002 814.5917 1202 2.054 0.035 2.289 0.051 1.114 0.0002 857.61861202 3.614 0.081 4.049 0.076 1.121 0.0002 458.2405 1101 1.632 0.0541.986 0.077 1.217 0.0003 481.3172 1202 3.866 0.050 4.159 0.060 1.0760.0003 541.3139 1101 4.022 0.081 4.494 0.096 1.117 0.0003 545.3453 11013.500 0.071 3.898 0.078 1.114 0.0003 592.3571 1202 2.113 0.095 2.5600.062 1.212 0.0003 757.5014 1204 3.642 0.067 3.971 0.052 1.091 0.0003833.7551 1204 2.061 0.223 3.140 0.161 1.524 0.0003 543.3296 1101 3.6520.055 4.023 0.087 1.102 0.0004 605.5457 1203 5.025 0.091 5.509 0.0891.096 0.0004 731.4916 1204 5.419 0.060 5.746 0.063 1.060 0.0004 775.55331202 2.106 0.063 2.449 0.068 1.163 0.0004 813.5885 1202 3.034 0.0433.276 0.048 1.080 0.0004 542.3173 1101 2.234 0.087 2.687 0.089 1.2030.0005 722.5335 1101 2.406 0.062 2.717 0.056 1.129 0.0005 744.55 12021.842 0.064 2.222 0.083 1.206 0.0005 769.5656 1204 3.889 0.055 4.1740.054 1.073 0.0005 807.59 1202 2.348 0.061 2.718 0.082 1.158 0.0005828.5743 1202 5.181 0.076 5.598 0.085 1.081 0.0005 521.4522 1203 2.1240.191 3.028 0.158 1.425 0.0007 604.5433 1203 6.440 0.091 6.901 0.0891.072 0.0007 744.4956 1204 3.851 0.049 4.117 0.058 1.069 0.0007 748.52871204 2.249 0.127 2.820 0.088 1.254 0.0007 771.5814 1204 4.118 0.0514.367 0.046 1.061 0.0007 827.5701 1202 6.245 0.082 6.684 0.092 1.0700.0007 747.5201 1202 1.886 0.083 2.313 0.088 1.226 0.0008 775.5528 12043.176 0.061 3.537 0.085 1.114 0.0008 787.5465 1204 2.543 0.116 3.0170.054 1.187 0.0008 827.5448 1101 3.373 0.104 3.873 0.095 1.148 0.0008546.3485 1101 1.890 0.073 2.287 0.089 1.210 0.0009 520.4499 1203 3.7750.176 4.577 0.148 1.212 0.001 570.3725 1202 2.926 0.047 3.155 0.0451.078 0.001 781.562 1201 7.334 0.054 7.638 0.071 1.041 0.001 632.50321203 1.435 0.112 0.795 0.154 0.554 0.0011 743.5469 1202 2.976 0.0703.348 0.086 1.125 0.0012 768.5539 1204 3.928 0.069 4.279 0.077 1.0890.0012 806.5873 1202 4.280 0.057 4.635 0.092 1.083 0.0012 575.2728 11012.419 0.061 2.739 0.074 1.132 0.0013 550.4957 1203 6.766 0.117 7.4150.160 1.096 0.0014 805.5839 1202 5.470 0.060 5.841 0.097 1.068 0.0014183.0661 1101 2.412 0.147 3.015 0.094 1.250 0.0015 551.4986 1203 5.3500.117 5.988 0.158 1.119 0.0015 741.5305 1204 2.825 0.080 3.250 0.1021.150 0.0015 541.3435 1202 5.595 0.072 5.974 0.094 1.068 0.0018 552.50221203 3.055 0.110 3.643 0.147 1.193 0.0018 506.3213 1202 2.487 0.0542.748 0.061 1.105 0.0019 569.369 1202 4.867 0.049 5.089 0.047 1.0460.0019 782.5653 1201 6.310 0.058 6.614 0.076 1.048 0.002 490.3641 12032.028 0.121 1.397 0.159 0.689 0.0021 542.3461 1202 3.803 0.061 4.1060.074 1.080 0.0021 566.3434 1202 5.130 0.071 5.436 0.062 1.060 0.0022833.7571 1203 2.853 0.122 3.507 0.170 1.229 0.0022 837.718 1204 2.5930.250 3.662 0.217 1.412 0.0023 549.4845 1203 5.510 0.107 5.994 0.1101.088 0.0025 793.5681 1204 3.116 0.058 3.372 0.055 1.082 0.0025 478.2541201 1.629 0.116 1.051 0.148 0.645 0.0027 536.4794 1203 2.188 0.1342.799 0.143 1.279 0.0027 548.4815 1203 6.949 0.103 7.405 0.103 1.0660.0027 832.6037 1202 4.981 0.069 5.274 0.062 1.059 0.0028 830.5894 12024.738 0.076 5.051 0.064 1.066 0.0029 1098.9739 1204 3.097 0.070 2.6060.152 0.842 0.0031 767.5495 1204 5.033 0.074 5.369 0.082 1.067 0.0034675.6377 1204 3.875 0.066 4.160 0.066 1.074 0.0035 564.5134 1203 2.6410.093 3.075 0.111 1.164 0.0036 789.5892 1202 1.703 0.053 1.952 0.0651.147 0.0036 832.7523 1203 3.734 0.131 4.374 0.169 1.171 0.0036 748.57351202 3.940 0.052 3.722 0.050 0.945 0.0037 801.555 1202 2.543 0.077 2.8810.082 1.133 0.0037 856.672 1202 2.777 0.048 2.553 0.057 0.919 0.0037858.6861 1202 2.982 0.055 2.756 0.052 0.924 0.0043 803.5677 1202 7.1010.095 7.502 0.098 1.057 0.0045 838.7226 1204 2.127 0.200 2.973 0.2061.398 0.0046 685.26 1202 1.708 0.074 1.998 0.063 1.169 0.0047 826.70691204 2.279 0.148 2.819 0.099 1.237 0.0047 582.2473 1201 3.332 0.1222.793 0.141 0.838 0.0049 590.343 1202 4.005 0.069 4.304 0.076 1.0750.005 832.7492 1204 3.163 0.146 3.848 0.190 1.217 0.0051 829.5859 12025.864 0.082 6.181 0.070 1.054 0.0053 772.5862 1204 3.263 0.043 3.4370.042 1.053 0.0054 409.0208 1202 2.753 0.050 2.980 0.063 1.082 0.0055509.3493 1202 2.358 0.054 2.579 0.055 1.094 0.0056 589.3403 1202 5.8470.076 6.171 0.085 1.056 0.0057 430.3818 1204 4.790 0.109 5.253 0.1221.097 0.0061 804.5718 1202 5.801 0.078 6.117 0.080 1.054 0.0066 505.32291202 3.931 0.071 4.222 0.076 1.074 0.0068 523.3634 1101 3.365 0.1113.791 0.103 1.127 0.0072 671.5723 1204 2.131 0.140 2.604 0.087 1.2220.0073 830.7352 1204 4.091 0.130 4.638 0.150 1.134 0.0073 431.386 12043.354 0.108 3.795 0.118 1.132 0.0074 631.628 1203 1.746 0.166 2.3910.161 1.369 0.0075 825.6926 1203 1.644 0.169 2.235 0.124 1.359 0.0079614.4914 1203 2.821 0.080 2.448 0.114 0.868 0.008 831.6001 1202 6.1090.077 6.404 0.074 1.048 0.0081 615.3539 1202 2.394 0.063 2.629 0.0581.098 0.0083 568.3573 1202 3.959 0.049 4.167 0.060 1.053 0.0088 767.5471202 2.992 0.097 3.358 0.092 1.122 0.0088 768.5503 1202 1.849 0.1252.277 0.090 1.231 0.0089 831.7408 1203 3.971 0.109 4.425 0.130 1.1140.0089 411.3212 1202 2.838 0.054 3.033 0.048 1.069 0.0099 796.5876 12042.633 0.059 2.860 0.062 1.087 0.0099 863.6876 1204 4.853 0.087 5.1880.092 1.069 0.0104 492.3816 1203 3.588 0.076 3.302 0.077 0.920 0.0105772.5278 1204 3.282 0.056 3.488 0.055 1.063 0.0107 825.5544 1202 2.6440.112 3.084 0.126 1.167 0.0108 320.2356 1204 1.302 0.114 1.686 0.0851.295 0.011 380.3096 1204 1.580 0.083 1.899 0.090 1.202 0.0112 519.33211101 3.914 0.122 4.382 0.133 1.119 0.0116 711.2577 1202 2.205 0.0752.454 0.056 1.113 0.0118 493.385 1203 2.083 0.073 1.722 0.124 0.8270.0119 565.3394 1202 6.924 0.073 7.186 0.068 1.038 0.0119 670.569 12043.179 0.090 3.515 0.093 1.106 0.012 856.6061 1202 4.500 0.081 4.8050.086 1.068 0.0121 340.2976 1203 1.502 0.124 1.937 0.112 1.290 0.0124287.2824 1101 1.958 0.180 1.265 0.202 0.646 0.0126 495.332 1101 5.1960.094 5.565 0.112 1.071 0.0137 304.241 1204 4.876 0.053 5.088 0.0661.044 0.0138 305.2438 1204 2.492 0.061 2.719 0.067 1.091 0.0145 616.50521203 4.260 0.086 3.957 0.084 0.929 0.0145 746.5119 1204 2.615 0.1653.137 0.119 1.200 0.0152 861.7806 1203 2.533 0.092 2.902 0.119 1.1460.0152 686.4877 1204 2.617 0.088 2.881 0.052 1.101 0.0155 830.7363 12034.834 0.111 5.263 0.135 1.089 0.0162 835.7006 1204 2.651 0.155 3.2140.167 1.212 0.0165 867.7579 1204 2.654 0.185 3.308 0.189 1.246 0.0167870.7307 1203 3.361 0.074 2.985 0.142 0.888 0.0169 246.1465 1202 3.7610.084 4.072 0.096 1.083 0.017 507.3316 1202 2.891 0.047 3.094 0.0711.070 0.0173 855.6023 1202 5.497 0.079 5.784 0.087 1.052 0.0173 578.52771203 4.183 0.420 5.548 0.355 1.326 0.0176 615.4938 1203 1.386 0.1330.902 0.148 0.651 0.0177 808.5803 1201 5.667 0.029 5.784 0.039 1.0210.0177 860.7752 1204 3.643 0.124 4.070 0.125 1.117 0.0185 518.4345 12031.584 0.180 2.199 0.178 1.388 0.0186 520.3354 1101 2.215 0.123 2.6460.130 1.195 0.0188 765.5313 1202 1.717 0.123 2.098 0.095 1.222 0.0195777.5287 1201 2.793 0.063 3.020 0.072 1.082 0.0198 887.8005 1203 2.9480.118 3.306 0.086 1.122 0.0198 739.5143 1204 2.602 0.149 3.121 0.1581.200 0.0199 831.7387 1204 3.193 0.121 3.648 0.150 1.142 0.02 521.34771101 3.643 0.099 4.021 0.127 1.104 0.0203 584.2646 1204 3.154 0.1302.649 0.172 0.840 0.0207 661.6233 1204 2.430 0.088 2.715 0.079 1.1170.0209 715.5167 1204 2.449 0.131 2.902 0.141 1.185 0.0218 529.3167 12023.031 0.071 3.265 0.069 1.077 0.0221 544.4481 1203 2.105 0.067 2.3330.071 1.108 0.0234 866.7532 1204 4.149 0.145 4.693 0.189 1.131 0.0235581.3344 1202 1.844 0.111 2.207 0.110 1.197 0.0245 795.5838 1204 3.5690.060 3.769 0.063 1.056 0.0246 807.5768 1201 6.720 0.030 6.829 0.0371.016 0.0256 854.5902 1202 3.801 0.070 4.031 0.073 1.060 0.0266 865.74871204 4.895 0.152 5.445 0.193 1.112 0.0268 496.3355 1101 3.396 0.0953.736 0.119 1.100 0.027 755.5467 1201 3.507 0.072 3.755 0.084 1.0710.0274 1019.3838 1102 3.035 0.036 2.919 0.037 0.962 0.0291 684.5489 12042.508 0.071 2.745 0.080 1.094 0.0294 774.0316 1204 1.789 0.192 1.1670.203 0.652 0.0294 765.5334 1204 3.218 0.101 3.540 0.104 1.100 0.0299630.5587 1203 3.126 0.072 3.344 0.064 1.070 0.0301 488.3873 1203 2.2410.069 1.986 0.096 0.886 0.0321 302.2255 1204 3.506 0.085 3.765 0.0811.074 0.0322 757.5625 1201 7.689 0.066 7.909 0.077 1.029 0.0325 617.50891203 2.978 0.089 2.698 0.092 0.906 0.0328 829.7239 1204 2.802 0.1203.185 0.131 1.137 0.0344 303.1079 1202 5.656 0.048 5.802 0.047 1.0260.0348 826.5581 1202 1.648 0.107 2.032 0.149 1.233 0.0362 506.2851 12013.080 0.117 2.656 0.165 0.862 0.0363 504.3814 1203 1.746 0.098 1.4130.124 0.809 0.0366 691.1957 1102 2.130 0.060 1.938 0.068 0.910 0.0373626.5278 1203 3.820 0.064 4.009 0.060 1.049 0.0375 522.3511 1101 1.7530.091 2.069 0.121 1.180 0.0385 759.5779 1201 6.294 0.076 6.541 0.0901.039 0.0386 625.5161 1203 2.856 0.057 3.045 0.070 1.066 0.0387 760.58111201 5.173 0.077 5.422 0.091 1.048 0.0395 484.3794 1204 2.181 0.0921.776 0.178 0.815 0.0396 819.5642 1202 2.811 0.080 3.104 0.120 1.1040.0404 853.5862 1202 4.775 0.069 4.985 0.075 1.044 0.0423 700.552 11012.278 0.124 2.617 0.102 1.149 0.0426 709.2594 1202 2.152 0.092 2.3820.053 1.107 0.0426 662.5175 1204 3.806 0.120 3.420 0.145 0.899 0.0432761.5846 1201 3.118 0.074 3.357 0.091 1.077 0.0436 743.5466 1203 2.0110.077 2.266 0.099 1.127 0.0442 478.3664 1203 1.545 0.088 1.212 0.1430.785 0.0447 784.5811 1201 5.585 0.073 5.809 0.082 1.040 0.0448 860.77561203 4.383 0.087 4.653 0.100 1.061 0.0448 601.5164 1203 7.642 0.0567.463 0.069 0.976 0.0453 758.5655 1201 6.637 0.074 6.860 0.084 1.0340.0488 371.3542 1203 3.253 0.055 3.435 0.074 1.056 0.0489 783.5778 12016.661 0.074 6.883 0.083 1.033 0.049 921.8145 1204 2.716 0.211 3.2550.155 1.198 0.0494 824.6892 1203 2.657 0.108 2.964 0.108 1.116 0.0497

TABLE 3 Accurate mass features differing between clinically diagnosed ADpatients with a significant cognitive impairment and clinicallydiagnosed non-AD patients with a significant cognitive impairment (p <0.05, log2 transformed). Detected Analysis AVG (log2) SEM AVG (log2) SEMlog(2) Mass Mode AD + Cog AD + Cog Non-AD + Cog Non-AD + Cog Ratio PValue 723.5197 1204 3.581 0.053 4.571 0.070 0.783 1.07E−17 723.5195 12022.193 0.056 3.033 0.049 0.723 2.70E−17 749.5367 1202 3.187 0.044 3.8120.045 0.836 5.25E−15 724.5257 1204 2.850 0.048 3.630 0.066 0.7851.68E−14 752.5564 1202 2.281 0.049 2.975 0.056 0.767 5.71E−14 751.55551204 4.591 0.056 5.427 0.071 0.846 6.78E−14 751.5529 1202 3.369 0.0524.055 0.054 0.831 1.46E−13 752.5583 1204 3.484 0.056 4.284 0.070 0.8132.38E−13 749.5407 1204 4.423 0.047 5.185 0.073 0.853 5.02E−13 750.54021202 2.210 0.052 2.804 0.044 0.788 9.50E−13 750.544 1204 3.258 0.0544.004 0.074 0.814 7.49E−12 725.5385 1204 2.815 0.058 3.543 0.078 0.7941.10E−10 699.5198 1204 2.129 0.055 2.825 0.079 0.754 3.41E−10 727.55681204 3.445 0.055 4.039 0.076 0.853 1.48E−08 728.5627 1204 2.875 0.0473.301 0.056 0.871 1.21E−07 726.5461 1204 2.759 0.044 3.175 0.057 0.8691.35E−07 775.5528 1204 3.176 0.061 3.712 0.075 0.856 3.75E−07 813.58851202 3.034 0.043 3.369 0.049 0.901 2.08E−06 775.5533 1202 2.106 0.0632.524 0.056 0.834 4.26E−06 747.5245 1204 3.433 0.058 3.933 0.088 0.8738.18E−06 814.5917 1202 2.054 0.035 2.355 0.055 0.872 1.16E−05 747.52011202 1.886 0.083 2.357 0.062 0.800 2.25E−05 811.5732 1202 2.729 0.0473.041 0.055 0.897 4.54E−05 789.5892 1202 1.703 0.053 2.013 0.061 0.8460.0002 810.5399 1204 2.760 0.071 3.111 0.064 0.887 0.0004 795.5838 12043.569 0.060 3.845 0.050 0.928 0.0007 856.6061 1202 4.500 0.081 4.8940.078 0.919 0.0008 783.5148 1204 3.217 0.063 3.512 0.061 0.916 0.0011855.6023 1202 5.497 0.079 5.876 0.079 0.935 0.0011 858.6212 1202 2.5330.080 2.917 0.080 0.868 0.0011 787.5465 1204 2.543 0.116 2.964 0.0450.858 0.0013 857.6186 1202 3.614 0.081 3.989 0.081 0.906 0.0015 773.5371202 1.581 0.066 1.875 0.065 0.843 0.002 784.5237 1204 3.211 0.056 3.4640.056 0.927 0.002 748.5287 1204 2.249 0.127 2.811 0.123 0.800 0.0021828.5743 1202 5.181 0.076 5.517 0.075 0.939 0.0023 827.5701 1202 6.2450.082 6.608 0.083 0.945 0.0026 786.5416 1204 3.729 0.049 3.961 0.0580.941 0.0029 871.5934 1202 1.841 0.085 2.189 0.076 0.841 0.0031 744.55361204 4.260 0.047 4.473 0.054 0.952 0.0037 787.5729 1202 1.834 0.0542.081 0.065 0.881 0.0042 755.486 1204 3.146 0.061 3.395 0.059 0.9270.0044 796.5876 1204 2.633 0.059 2.868 0.056 0.918 0.0048 812.5762 12021.752 0.045 2.034 0.088 0.861 0.0049 817.5377 1102 2.175 0.064 2.4270.060 0.896 0.0053 383.3284 1204 1.263 0.120 1.722 0.107 0.733 0.0055840.6063 1202 2.746 0.042 2.927 0.050 0.938 0.0065 544.397 1204 2.9980.111 2.235 0.255 1.342 0.0066 570.3725 1202 2.926 0.047 3.115 0.0490.939 0.0071 782.5085 1204 3.369 0.056 3.596 0.062 0.937 0.0077 847.59541202 2.317 0.089 2.662 0.090 0.870 0.0079 855.6016 1102 3.386 0.0603.619 0.062 0.936 0.0082 769.5656 1204 3.889 0.055 4.103 0.057 0.9480.0084 819.5642 1202 2.811 0.080 3.149 0.099 0.893 0.0089 828.5737 11022.982 0.056 3.197 0.058 0.933 0.009 590.343 1202 4.005 0.069 4.261 0.0660.940 0.0091 719.6231 1204 4.528 0.074 4.158 0.119 1.089 0.0092 589.34031202 5.847 0.076 6.129 0.075 0.954 0.0098 768.5539 1204 3.928 0.0694.189 0.071 0.938 0.0102 839.6031 1202 3.723 0.041 3.895 0.051 0.9560.0102 821.5712 1102 2.919 0.053 3.120 0.056 0.936 0.0106 1226.097 12033.278 0.092 2.921 0.105 1.122 0.0122 664.5323 1204 4.909 0.126 4.3910.161 1.118 0.0126 733.6426 1204 2.981 0.074 2.625 0.120 1.135 0.0126771.5814 1204 4.118 0.051 4.313 0.057 0.955 0.0126 809.5937 1201 6.4530.060 6.677 0.065 0.966 0.0126 720.6258 1204 3.565 0.076 3.215 0.1171.109 0.0127 667.5475 1204 3.516 0.124 3.013 0.157 1.167 0.0132 665.53541204 3.705 0.121 3.182 0.170 1.164 0.0133 666.5456 1204 4.713 0.1264.232 0.146 1.114 0.0142 793.5386 1102 3.435 0.052 3.632 0.061 0.9460.015 810.5971 1201 5.436 0.063 5.665 0.068 0.959 0.015 832.6037 12024.981 0.069 5.231 0.074 0.952 0.0151 663.5216 1204 2.566 0.145 1.9460.210 1.319 0.0159 743.5471 1204 5.227 0.047 5.417 0.062 0.965 0.0162767.5495 1204 5.033 0.074 5.287 0.074 0.952 0.017 796.5292 1204 2.9280.063 3.142 0.062 0.932 0.017 671.5723 1204 2.131 0.140 2.568 0.1130.830 0.0173 758.5092 1204 4.504 0.047 4.670 0.051 0.964 0.0188 509.34931202 2.358 0.054 2.542 0.056 0.927 0.0192 827.57 1102 3.984 0.059 4.1910.064 0.951 0.0194 569.369 1202 4.867 0.049 5.033 0.049 0.967 0.02831.6001 1202 6.109 0.077 6.380 0.085 0.958 0.02 638.5149 1204 3.5300.120 3.086 0.150 1.144 0.0224 768.5503 1202 1.849 0.125 2.190 0.0800.845 0.0249 313.1153 1204 3.859 0.102 3.338 0.209 1.156 0.025 592.35711202 2.113 0.095 2.375 0.065 0.890 0.0262 794.5421 1102 2.364 0.0482.536 0.061 0.932 0.0276 662.5175 1204 3.806 0.120 3.377 0.153 1.1270.0292 670.569 1204 3.179 0.090 3.474 0.101 0.915 0.0312 870.7307 12033.361 0.074 3.012 0.144 1.116 0.0312 1098.974 1204 3.097 0.070 2.7390.151 1.131 0.0313 694.6161 1204 2.845 0.080 2.437 0.173 1.168 0.0324541.3432 1102 3.138 0.034 3.255 0.043 0.964 0.0335 581.3344 1202 1.8440.111 2.147 0.087 0.859 0.0351 691.596 1204 2.394 0.098 1.999 0.1591.198 0.0351 803.5677 1202 7.101 0.095 7.405 0.108 0.959 0.0364 591.35421202 3.870 0.092 4.110 0.064 0.942 0.0367 705.6093 1204 3.137 0.0722.823 0.132 1.111 0.0367 847.5316 1101 2.564 0.060 2.815 0.104 0.9110.0367 854.5902 1202 3.801 0.070 4.027 0.082 0.944 0.0376 215.9153 11014.405 0.297 5.221 0.247 0.844 0.0379 569.3687 1102 2.063 0.042 2.2170.062 0.931 0.0404 640.5294 1204 3.725 0.127 3.313 0.154 1.124 0.0407853.5862 1202 4.775 0.069 4.993 0.081 0.956 0.0413 830.5894 1202 4.7380.076 4.968 0.082 0.954 0.0415 760.5216 1204 4.008 0.048 4.156 0.0530.964 0.0417 530.3821 1204 2.614 0.172 2.016 0.240 1.296 0.0441 1225.0921203 3.451 0.135 3.011 0.170 1.146 0.0445 819.5551 1102 2.198 0.0582.369 0.062 0.928 0.0456 743.5469 1202 2.976 0.070 3.200 0.087 0.9300.0464 444.2717 1202 1.304 0.083 1.577 0.109 0.827 0.0471 678.5477 12043.874 0.101 3.523 0.145 1.099 0.0485 741.5305 1204 2.825 0.080 3.0860.104 0.916 0.0489 759.5163 1204 4.645 0.061 4.811 0.057 0.966 0.0493820.568 1202 1.757 0.128 2.102 0.117 0.836 0.0497

TABLE 4 Accurate mass features differing between clinically diagnosed ADpatients with a significant cognitive impairment and clinicallydiagnosed AD patients without a significant cognitive impairment (p <0.05, log2 transformed). Detected Analysis AVG (log2) SEM AVG (log2) SEMlog(2) Mass Mode AD AD AD noCog AD noCog Ratio P Value 541.3432 11023.138 0.034 3.518 0.039 0.892 3.47E−10 569.3687 1102 2.063 0.042 2.4790.045 0.832 3.90E−09 829.5856 1102 3.806 0.059 4.301 0.048 0.8851.30E−08 567.3547 1102 2.629 0.050 3.031 0.045 0.868 1.02E−07 831.59971102 4.372 0.055 4.774 0.042 0.916 1.93E−07 832.6026 1102 3.268 0.0553.662 0.042 0.892 2.67E−07 804.5713 1102 4.017 0.048 4.426 0.054 0.9083.40E−07 803.568 1102 5.238 0.048 5.656 0.057 0.926 3.97E−07 871.55281102 2.885 0.049 3.279 0.054 0.880 8.30E−07 565.3394 1102 3.265 0.0583.728 0.064 0.876 1.14E−06 805.5832 1102 3.874 0.056 4.292 0.055 0.9031.41E−06 819.5551 1102 2.198 0.058 2.604 0.053 0.844 2.52E−06 555.31021102 1.634 0.053 2.033 0.059 0.803 3.32E−06 853.5854 1102 2.449 0.0562.824 0.051 0.867 5.09E−06 805.5605 1101 5.184 0.074 5.665 0.065 0.9156.42E−06 808.5792 1101 4.488 0.071 4.925 0.057 0.911 9.48E−06 793.53861102 3.435 0.052 3.799 0.057 0.904 1.17E−05 807.5758 1101 5.531 0.0695.961 0.059 0.928 1.20E−05 806.5639 1101 4.199 0.075 4.669 0.066 0.8991.28E−05 837.5027 1101 3.402 0.050 3.781 0.064 0.900 1.45E−05 794.54211102 2.364 0.048 2.703 0.059 0.875 2.79E−05 780.5474 1101 5.241 0.0645.661 0.070 0.926 3.62E−05 779.5444 1101 6.239 0.065 6.654 0.070 0.9384.37E−05 821.5712 1102 2.919 0.053 3.221 0.048 0.906 0.0001 827.57 11023.984 0.059 4.337 0.063 0.919 0.0001 886.5582 1102 2.911 0.041 3.1890.053 0.913 0.0001 915.5191 1101 2.189 0.058 2.592 0.073 0.845 0.0001722.5335 1101 2.406 0.062 2.735 0.057 0.880 0.0002 828.5737 1102 2.9820.056 3.313 0.063 0.900 0.0002 833.5932 1101 3.045 0.095 3.522 0.0770.865 0.0002 855.6016 1102 3.386 0.060 3.704 0.055 0.914 0.0002 755.54681101 2.115 0.086 2.563 0.082 0.825 0.0003 795.555 1102 2.520 0.054 2.8180.057 0.894 0.0003 803.5445 1101 4.941 0.074 5.324 0.070 0.928 0.0003811.6096 1101 2.801 0.116 3.370 0.096 0.831 0.0003 804.5476 1101 3.9870.073 4.370 0.073 0.912 0.0004 829.5604 1101 3.360 0.075 3.728 0.0630.901 0.0004 783.5672 1101 3.535 0.092 3.993 0.085 0.885 0.0005 832.57911101 3.184 0.076 3.541 0.067 0.899 0.0008 847.5316 1101 2.564 0.0602.955 0.095 0.868 0.0008 784.5811 1101 4.161 0.112 4.668 0.093 0.8910.0009 831.5759 1101 4.123 0.080 4.485 0.066 0.919 0.0009 757.4991 11013.450 0.089 3.866 0.083 0.892 0.0011 781.5617 1101 5.928 0.084 6.3050.075 0.940 0.0014 795.5181 1101 2.458 0.082 2.823 0.073 0.871 0.0015782.565 1101 4.901 0.088 5.279 0.080 0.928 0.0022 760.5811 1101 4.2160.099 4.632 0.089 0.910 0.0027 759.5779 1101 5.352 0.096 5.752 0.0850.930 0.0028 591.3542 1202 3.870 0.092 4.224 0.068 0.916 0.0031 592.35711202 2.113 0.095 2.472 0.070 0.855 0.0035 761.5843 1101 2.241 0.0842.609 0.088 0.859 0.0035 858.6212 1202 2.533 0.080 2.870 0.078 0.8830.0037 458.2405 1101 1.632 0.054 1.858 0.054 0.878 0.0042 785.5933 11015.687 0.112 6.096 0.083 0.933 0.0047 786.5967 1101 4.611 0.112 5.0180.084 0.919 0.005 758.5656 1101 5.511 0.107 5.915 0.094 0.932 0.0061306.2569 1204 2.878 0.070 3.125 0.053 0.921 0.0066 857.6186 1202 3.6140.081 3.925 0.076 0.921 0.0066 757.5626 1101 6.470 0.106 6.861 0.0940.943 0.0075 809.5936 1101 4.786 0.104 5.150 0.081 0.929 0.0077 810.59691101 3.779 0.108 4.144 0.083 0.912 0.0095 807.59 1202 2.348 0.061 2.5740.062 0.912 0.0113 383.3284 1204 1.263 0.120 1.659 0.095 0.761 0.012786.5416 1204 3.729 0.049 3.903 0.046 0.955 0.012 744.55 1202 1.8420.064 2.093 0.074 0.880 0.0124 827.5448 1101 3.373 0.104 3.751 0.1050.899 0.0128 784.5237 1204 3.211 0.056 3.405 0.052 0.943 0.0133 746.51191204 2.615 0.165 3.058 0.057 0.855 0.0145 817.5377 1102 2.175 0.0642.405 0.066 0.905 0.0155 699.5198 1204 2.129 0.055 2.309 0.047 0.9220.0161 757.5014 1204 3.642 0.067 3.869 0.063 0.941 0.0161 826.7069 12042.279 0.148 2.684 0.086 0.849 0.0215 758.5092 1204 4.504 0.047 4.6520.043 0.968 0.0235 546.3485 1101 1.890 0.073 2.115 0.064 0.894 0.0236801.555 1202 2.543 0.077 2.788 0.077 0.912 0.0272 829.7239 1204 2.8020.120 3.161 0.105 0.886 0.0272 748.5721 1102 3.795 0.067 3.996 0.0600.950 0.0288 518.3174 1101 3.358 0.051 3.523 0.054 0.953 0.0296 826.55811202 1.648 0.107 1.983 0.108 0.831 0.0309 830.5894 1202 4.738 0.0764.956 0.064 0.956 0.0317 430.3818 1204 4.790 0.109 5.108 0.096 0.9380.0322 313.1153 1204 3.859 0.102 3.311 0.233 1.166 0.0327 755.5467 12013.507 0.072 3.728 0.072 0.941 0.0327 545.3453 1101 3.500 0.071 3.7130.069 0.943 0.0353 320.2356 1204 1.302 0.114 1.590 0.070 0.819 0.0363183.0661 1101 2.412 0.147 2.779 0.096 0.868 0.0416 825.5544 1202 2.6440.112 2.968 0.109 0.891 0.0418 549.4845 1203 5.510 0.107 5.818 0.1030.947 0.0424 829.5859 1202 5.864 0.082 6.087 0.070 0.963 0.0433 431.3861204 3.354 0.108 3.651 0.099 0.919 0.0463 784.5811 1201 5.585 0.0735.782 0.064 0.966 0.0463 777.5287 1201 2.793 0.063 2.986 0.072 0.9350.0466 278.2254 1204 4.828 0.056 4.996 0.062 0.966 0.0467 548.4815 12036.949 0.103 7.233 0.096 0.961 0.0472 777.553 1202 1.720 0.100 2.0020.098 0.859 0.0483 517.314 1101 5.399 0.050 5.547 0.055 0.973 0.0488661.6233 1204 2.430 0.088 2.638 0.054 0.921 0.0489

TABLE 5 Accurate mass features differing between clinically diagnosednon-AD patients and non-demented controls (p < 0.05, log2 transformed).Detected Analysis AVG (log2) SEM AVG (log2) SEM log(2) Mass Mode NON-ADNON-AD Normal Normal Ratio P Value 541.3432 1102 3.255 0.043 3.679 0.0480.885 3.43E−12 567.3547 1102 2.621 0.052 3.117 0.054 0.841 2.78E−11805.5605 1101 5.159 0.072 5.604 0.048 0.921 1.08E−09 806.5639 1101 4.1500.073 4.630 0.049 0.896 1.51E−09 804.5713 1102 4.159 0.056 4.466 0.0330.931 3.79E−09 783.5672 1101 3.515 0.077 4.126 0.079 0.852 4.13E−09781.5617 1101 5.919 0.072 6.403 0.072 0.924 4.19E−09 780.5474 1101 5.2760.075 5.691 0.043 0.927 4.35E−09 803.568 1102 5.379 0.056 5.656 0.0350.951 4.43E−09 779.5444 1101 6.291 0.073 6.659 0.043 0.945 5.24E−09760.5811 1101 4.034 0.079 4.733 0.103 0.852 6.28E−09 805.5832 1102 3.8970.057 4.344 0.068 0.897 6.49E−09 759.5779 1101 5.166 0.082 5.810 0.0990.889 1.04E−08 782.565 1101 4.878 0.077 5.415 0.078 0.901 1.14E−08829.5856 1102 3.952 0.051 4.261 0.046 0.927 1.44E−08 569.3687 1102 2.2170.062 2.639 0.048 0.840 1.99E−08 755.5468 1101 1.974 0.089 2.667 0.0850.740 2.63E−08 757.4991 1101 3.368 0.086 3.945 0.070 0.854 2.69E−08871.5528 1102 2.960 0.059 3.301 0.040 0.897 4.58E−08 804.5476 1101 4.1250.073 4.556 0.060 0.906 6.76E−08 803.5445 1101 5.084 0.072 5.478 0.0590.928 7.46E−08 758.5656 1101 5.440 0.089 5.968 0.078 0.912 8.14E−08565.3394 1102 3.359 0.081 3.834 0.055 0.876 8.59E−08 757.5626 1101 6.4170.087 6.886 0.079 0.932 1.74E−07 808.5792 1101 4.527 0.078 4.904 0.0470.923 2.30E−07 795.5181 1101 2.491 0.071 2.948 0.064 0.845 2.65E−07807.5758 1101 5.569 0.080 5.911 0.047 0.942 3.99E−07 731.5464 1101 1.4600.132 2.519 0.157 0.580 4.14E−07 837.5027 1101 3.437 0.071 3.810 0.0500.902 4.34E−07 761.5843 1101 2.053 0.092 2.740 0.106 0.749 5.01E−07784.5811 1101 4.092 0.095 4.666 0.089 0.877 5.41E−07 831.5997 1102 4.5120.057 4.749 0.040 0.950 6.07E−07 832.6026 1102 3.409 0.056 3.677 0.0400.927 6.72E−07 555.3102 1102 1.792 0.066 2.170 0.046 0.826 8.11E−07833.5932 1101 3.038 0.087 3.495 0.056 0.869 1.09E−06 829.5604 1101 3.3810.077 3.754 0.058 0.901 3.91E−06 811.6096 1101 2.826 0.097 3.401 0.0960.831 4.19E−06 785.5933 1101 5.629 0.100 6.075 0.080 0.927 5.08E−06786.5967 1101 4.566 0.101 5.041 0.078 0.906 5.43E−06 827.57 1102 4.1910.064 4.486 0.062 0.934 6.39E−06 828.5737 1102 3.197 0.058 3.478 0.0610.919 1.44E−05 810.5969 1101 3.785 0.100 4.234 0.080 0.894 2.51E−05809.5936 1101 4.804 0.094 5.194 0.079 0.925 2.81E−05 517.314 1101 5.3390.065 5.575 0.069 0.958 3.58E−05 495.332 1101 5.003 0.083 5.391 0.1120.928 0.0001 518.3174 1101 3.325 0.062 3.601 0.073 0.923 0.0001 793.53861102 3.632 0.061 3.827 0.039 0.949 0.0001 794.5421 1102 2.536 0.0612.763 0.040 0.918 0.0001 832.5791 1101 3.305 0.080 3.607 0.061 0.9160.0001 853.5854 1102 2.527 0.065 2.790 0.052 0.906 0.0001 915.5191 11012.278 0.083 2.632 0.063 0.866 0.0001 183.0661 1101 2.324 0.143 2.9200.094 0.796 0.0002 521.3477 1101 3.406 0.099 3.895 0.127 0.875 0.0002523.3634 1101 3.221 0.102 3.673 0.103 0.877 0.0002 543.3296 1101 3.5790.072 3.898 0.087 0.918 0.0002 795.555 1102 2.608 0.069 2.893 0.0650.902 0.0002 831.5759 1101 4.272 0.080 4.531 0.060 0.943 0.0002 886.55821102 2.998 0.049 3.149 0.041 0.952 0.0002 496.3355 1101 3.200 0.0863.620 0.119 0.884 0.0004 827.5448 1101 3.340 0.105 3.752 0.095 0.8900.0004 819.5551 1102 2.369 0.062 2.571 0.046 0.922 0.0005 522.3511 11011.480 0.111 2.005 0.121 0.738 0.0006 817.5377 1102 2.427 0.060 2.6270.057 0.924 0.001 545.3453 1101 3.512 0.082 3.776 0.078 0.930 0.0011575.2728 1101 2.381 0.076 2.653 0.074 0.897 0.0012 700.552 1101 2.1500.094 2.535 0.102 0.848 0.0012 722.5335 1101 2.437 0.062 2.632 0.0560.926 0.0014 520.3354 1101 2.075 0.117 2.564 0.130 0.809 0.0016 458.24051101 1.664 0.064 1.924 0.077 0.865 0.0017 542.3173 1101 2.268 0.0922.603 0.089 0.871 0.0017 519.3321 1101 3.807 0.121 4.245 0.133 0.8970.0019 546.3485 1101 1.931 0.069 2.216 0.089 0.872 0.0021 541.3139 11014.077 0.092 4.353 0.096 0.937 0.0025 1019.384 1102 3.095 0.042 2.8280.037 1.094 0.0026 804.7227 1203 1.612 0.233 2.530 0.220 0.637 0.0027831.7408 1203 3.839 0.137 4.286 0.130 0.896 0.0028 303.1079 1202 5.5750.057 5.621 0.047 0.992 0.0032 549.4845 1203 5.480 0.127 5.807 0.1100.944 0.0033 548.4815 1203 6.917 0.122 7.174 0.103 0.964 0.0034 746.57171204 3.115 0.058 3.257 0.061 0.957 0.0046 830.7363 1203 4.694 0.1395.098 0.135 0.921 0.0046 306.2569 1204 3.015 0.059 3.164 0.067 0.9530.0058 760.5216 1204 4.156 0.053 4.211 0.039 0.987 0.0058 732.4938 12044.274 0.060 4.370 0.057 0.978 0.0059 855.6016 1102 3.619 0.062 3.7520.065 0.965 0.0059 523.4679 1203 3.249 0.212 3.886 0.165 0.836 0.0065522.4635 1203 4.717 0.208 5.313 0.176 0.888 0.0066 1227.107 1203 3.2350.116 3.547 0.100 0.912 0.0074 833.7551 1204 2.396 0.213 3.042 0.1610.788 0.0075 520.4499 1203 3.898 0.194 4.434 0.148 0.879 0.0076 1228.1111203 2.587 0.203 3.131 0.105 0.826 0.0077 521.4522 1203 2.319 0.2042.933 0.158 0.790 0.0083 591.3542 1202 4.110 0.064 4.209 0.058 0.9760.0085 856.672 1202 2.741 0.042 2.473 0.057 1.108 0.0086 777.553 12021.958 0.096 2.309 0.130 0.848 0.009 552.5022 1203 3.097 0.142 3.5290.147 0.878 0.0092 777.5287 1201 2.781 0.055 2.926 0.072 0.951 0.0092675.6377 1204 3.806 0.112 4.030 0.066 0.945 0.0098 748.5735 1202 3.8830.036 3.606 0.050 1.077 0.0101 467.807 1101 1.408 0.134 0.863 0.1441.631 0.0104 832.7523 1203 3.763 0.160 4.237 0.169 0.888 0.0104 837.7181204 2.737 0.273 3.548 0.217 0.771 0.0107 745.5658 1204 4.005 0.0604.109 0.068 0.975 0.0109 568.3573 1202 3.960 0.053 4.037 0.060 0.9810.0113 550.4957 1203 6.848 0.150 7.184 0.160 0.953 0.0114 833.7571 12032.911 0.156 3.397 0.170 0.857 0.0115 838.7226 1204 2.158 0.237 2.8800.206 0.749 0.0119 551.4986 1203 5.429 0.150 5.801 0.158 0.936 0.0121585.2673 1202 1.612 0.153 1.034 0.148 1.558 0.0125 821.5712 1102 3.1200.056 3.200 0.044 0.975 0.0133 835.7006 1204 2.553 0.198 3.113 0.1670.820 0.0136 731.4916 1204 5.523 0.062 5.567 0.063 0.992 0.0138 867.75791204 2.503 0.252 3.205 0.189 0.781 0.0139 482.3215 1202 2.040 0.0582.181 0.062 0.935 0.0145 858.6861 1202 2.935 0.050 2.670 0.052 1.0990.0145 584.2646 1204 3.282 0.185 2.567 0.172 1.279 0.0148 829.7246 12034.743 0.119 4.973 0.101 0.954 0.015 828.7207 1203 5.497 0.121 5.7160.107 0.962 0.0154 723.5197 1204 4.571 0.070 4.214 0.056 1.085 0.0167864.738 1204 4.725 0.142 5.051 0.141 0.936 0.0171 691.1957 1102 2.1680.067 1.878 0.068 1.155 0.0181 585.2664 1204 1.716 0.183 1.077 0.1701.593 0.0187 749.5777 1202 2.879 0.034 2.659 0.046 1.083 0.0189 757.50141204 3.770 0.066 3.847 0.052 0.980 0.0197 863.7336 1204 5.458 0.1465.763 0.147 0.947 0.02 830.7352 1204 4.133 0.151 4.493 0.150 0.9200.0202 252.2095 1204 1.665 0.076 1.852 0.070 0.899 0.0206 860.7756 12034.306 0.109 4.507 0.100 0.955 0.0222 863.6876 1204 4.889 0.090 5.0260.092 0.973 0.0227 861.7806 1203 2.435 0.158 2.811 0.119 0.866 0.0228840.6063 1202 2.927 0.050 2.664 0.059 1.099 0.0229 632.5762 1203 1.3250.170 1.773 0.132 0.747 0.0231 826.7069 1204 2.424 0.137 2.731 0.0990.887 0.0242 419.8773 1101 1.946 0.176 1.305 0.194 1.491 0.0243 440.35321204 1.481 0.134 0.945 0.180 1.567 0.0247 831.7387 1204 3.137 0.1643.534 0.150 0.888 0.0248 310.2883 1204 2.448 0.085 2.607 0.062 0.9390.0256 481.3172 1202 3.978 0.053 4.029 0.060 0.987 0.0259 205.8865 11013.759 0.234 2.793 0.319 1.346 0.0271 513.4122 1204 1.142 0.153 0.6230.163 1.833 0.0279 839.6031 1202 3.895 0.051 3.609 0.057 1.079 0.0286865.7487 1204 4.847 0.188 5.275 0.193 0.919 0.029 579.5325 1203 4.1880.154 4.491 0.127 0.933 0.0292 807.59 1202 2.477 0.072 2.633 0.082 0.9410.0293 351.8903 1101 2.359 0.220 1.567 0.256 1.506 0.0296 782.5085 12043.596 0.062 3.662 0.055 0.982 0.0297 859.7696 1203 5.853 0.104 5.9720.093 0.980 0.0298 409.8484 1101 2.107 0.192 1.437 0.210 1.466 0.0304577.5137 1203 8.099 0.104 8.148 0.094 0.994 0.0305 1018.942 1203 3.4680.142 3.767 0.126 0.921 0.0305 703.5709 1101 3.024 0.090 3.185 0.0790.949 0.0314 484.3794 1204 2.234 0.118 1.721 0.178 1.298 0.0315 283.26021204 6.622 0.070 6.621 0.067 1.000 0.0316 401.8166 1101 1.696 0.1611.139 0.177 1.489 0.0316 832.7492 1204 3.251 0.195 3.728 0.190 0.8720.0316 190.033 1101 3.264 0.233 2.384 0.289 1.369 0.0317 806.5873 12024.367 0.082 4.490 0.092 0.973 0.032 282.2573 1204 9.088 0.073 9.0190.071 1.008 0.0327 195.8577 1101 3.161 0.202 2.370 0.266 1.334 0.0328744.4956 1204 3.941 0.058 3.988 0.058 0.988 0.0332 866.7532 1204 4.1230.184 4.547 0.189 0.907 0.0334 215.9153 1101 5.221 0.247 4.183 0.3461.248 0.0335 825.6926 1203 1.767 0.173 2.165 0.124 0.816 0.0339 469.80421101 1.617 0.150 1.096 0.170 1.475 0.0344 874.7645 1204 3.091 0.1443.377 0.111 0.915 0.0345 494.4343 1203 2.119 0.275 2.809 0.232 0.7540.0346 399.8196 1101 1.827 0.171 1.249 0.184 1.463 0.0348 802.7056 12041.701 0.206 2.248 0.202 0.756 0.0348 576.5098 1203 9.446 0.111 9.4600.099 0.999 0.0355 382.1082 1101 2.222 0.229 1.494 0.220 1.487 0.0356720.6438 1204 2.962 0.180 3.293 0.083 0.900 0.036 805.5839 1202 5.5740.081 5.658 0.097 0.985 0.0365 779.5441 1201 7.096 0.041 7.010 0.0531.012 0.0368 231.8893 1101 2.592 0.233 1.800 0.259 1.440 0.0372 686.48771204 2.721 0.054 2.791 0.052 0.975 0.0374 429.3749 1204 2.601 0.0982.777 0.077 0.937 0.0378 265.8423 1101 2.499 0.220 1.743 0.252 1.4330.0384 722.6424 1204 3.726 0.167 4.006 0.086 0.930 0.0385 780.5474 12016.022 0.043 5.972 0.054 1.008 0.0396 599.4936 1204 1.373 0.191 1.8730.188 0.733 0.04 536.4794 1203 2.280 0.202 2.711 0.143 0.841 0.0422244.0559 1101 3.574 0.126 3.098 0.132 1.153 0.0424 202.0453 1101 5.5610.180 4.832 0.215 1.151 0.0425 535.7944 1101 1.480 0.143 1.018 0.1531.455 0.0425 758.5092 1204 4.670 0.051 4.658 0.042 1.003 0.0426 626.51041204 2.100 0.135 2.391 0.114 0.878 0.0427 847.5316 1101 2.815 0.1042.994 0.083 0.940 0.0432 592.3571 1202 2.375 0.065 2.480 0.062 0.9580.0437 569.4784 1204 1.175 0.183 0.634 0.178 1.853 0.0447 721.639 12044.879 0.142 5.064 0.088 0.963 0.0447 752.5583 1204 4.284 0.070 3.9660.061 1.080 0.0451 507.7055 1101 1.487 0.126 1.052 0.154 1.413 0.0454827.7086 1203 3.916 0.119 4.098 0.096 0.956 0.046 784.5237 1204 3.4640.056 3.490 0.037 0.993 0.0461 755.486 1204 3.395 0.059 3.450 0.0570.984 0.0471 628.5237 1204 2.627 0.158 2.955 0.136 0.889 0.0472 218.01921101 3.591 0.258 2.716 0.302 1.322 0.049

TABLE 6 Accurate mass features differing between clinically diagnosed ADpatients with a mild cognitive impairment and non-demented controls (p <0.05, log2 transformed). AVG AVG Detected Analysis (log2) SEM (log2) SEMMass Mode AD noCog AD noCog Normal Normal log(2) Ratio P Value 723.51971204 3.576 0.058 4.350 0.056 0.822 6.42E−14 723.5195 1202 2.181 0.0562.892 0.052 0.754 2.49E−13 751.5529 1202 3.293 0.050 3.920 0.050 0.8401.39E−12 724.5257 1204 2.883 0.044 3.463 0.055 0.833 9.96E−12 749.53671202 3.168 0.051 3.714 0.041 0.853 1.33E−11 752.5564 1202 2.216 0.0572.836 0.050 0.781 1.91E−11 751.5555 1204 4.553 0.060 5.248 0.060 0.8681.96E−11 750.5402 1202 2.175 0.054 2.704 0.044 0.804 2.92E−10 752.55831204 3.453 0.063 4.094 0.061 0.844 6.11E−10 750.544 1204 3.299 0.0523.858 0.063 0.855 3.31E−09 749.5407 1204 4.429 0.061 5.012 0.064 0.8849.50E−09 699.5198 1204 2.309 0.047 2.775 0.054 0.832 1.20E−08 725.53851204 2.958 0.060 3.417 0.054 0.866 4.59E−07 727.5568 1204 3.588 0.0503.986 0.060 0.900 3.00E−06 812.5762 1202 1.571 0.087 2.084 0.058 0.7541.13E−05 541.3432 1102 3.518 0.039 3.798 0.048 0.926 2.42E−05 813.58851202 2.992 0.041 3.276 0.048 0.913 2.68E−05 811.5732 1202 2.683 0.0553.027 0.055 0.886 3.92E−05 726.5461 1204 2.869 0.047 3.150 0.050 0.9110.0001 814.5917 1202 2.028 0.040 2.289 0.051 0.886 0.0001 632.5762 12030.944 0.175 1.830 0.132 0.516 0.0002 728.5627 1204 2.998 0.045 3.2810.060 0.914 0.0003 782.5085 1204 3.435 0.071 3.780 0.055 0.909 0.0003569.3687 1102 2.479 0.045 2.724 0.048 0.910 0.0004 803.5445 1101 5.3240.070 5.655 0.059 0.941 0.0007 804.5476 1101 4.370 0.073 4.703 0.0600.929 0.0009 817.5377 1102 2.405 0.066 2.712 0.057 0.887 0.001 787.57291202 1.864 0.058 2.145 0.057 0.869 0.0011 747.5245 1204 3.521 0.0633.886 0.090 0.906 0.0013 775.5533 1202 2.140 0.062 2.449 0.068 0.8740.0013 746.5717 1204 3.117 0.043 3.362 0.061 0.927 0.0014 827.57 11024.337 0.063 4.630 0.062 0.937 0.0017 745.5658 1204 3.989 0.041 4.2420.068 0.940 0.0018 828.5737 1102 3.313 0.063 3.590 0.061 0.923 0.0025542.3173 1101 2.344 0.067 2.687 0.089 0.872 0.0027 760.5216 1204 4.1400.052 4.347 0.039 0.952 0.0028 748.5287 1204 2.319 0.131 2.820 0.0880.822 0.003 774.0316 1204 2.044 0.199 1.167 0.203 1.752 0.0031 784.52371204 3.405 0.052 3.603 0.037 0.945 0.0035 775.5528 1204 3.221 0.0653.537 0.085 0.911 0.004 744.5536 1204 4.388 0.046 4.605 0.058 0.9530.0042 482.3215 1202 2.001 0.057 2.251 0.062 0.889 0.0043 768.5539 12043.995 0.060 4.279 0.077 0.934 0.0044 506.2851 1201 3.210 0.100 2.6560.165 1.209 0.0045 747.5201 1202 2.006 0.060 2.313 0.088 0.867 0.0045755.486 1204 3.295 0.069 3.561 0.057 0.925 0.0047 781.5617 1101 6.3050.075 6.610 0.072 0.954 0.005 786.5416 1204 3.903 0.046 4.087 0.0430.955 0.0052 804.5713 1102 4.426 0.054 4.610 0.033 0.960 0.0064 743.54711204 5.349 0.054 5.579 0.063 0.959 0.0066 541.3139 1101 4.172 0.0674.494 0.096 0.928 0.0069 767.5495 1204 5.085 0.063 5.369 0.082 0.9470.0072 782.565 1101 5.279 0.080 5.589 0.078 0.945 0.0074 555.3102 11022.033 0.059 2.240 0.046 0.908 0.0086 567.3547 1102 3.031 0.045 3.2180.054 0.942 0.0089 565.3394 1102 3.728 0.064 3.958 0.055 0.942 0.0094582.2473 1201 3.346 0.151 2.793 0.141 1.198 0.0099 803.568 1102 5.6560.057 5.838 0.035 0.969 0.0108 779.5444 1101 6.654 0.070 6.874 0.0430.968 0.0113 796.5876 1204 2.630 0.063 2.860 0.062 0.919 0.0117 758.50921204 4.652 0.043 4.808 0.042 0.968 0.0126 783.5148 1204 3.344 0.0683.574 0.056 0.936 0.0126 748.5735 1202 3.893 0.045 3.722 0.050 1.0460.0135 529.3167 1202 3.028 0.064 3.265 0.069 0.927 0.0138 781.562 12017.406 0.060 7.638 0.071 0.970 0.0144 780.5474 1101 5.661 0.070 5.8750.043 0.964 0.0145 810.5399 1204 2.905 0.069 3.145 0.066 0.923 0.0147837.5881 1202 2.412 0.040 2.577 0.053 0.936 0.0147 590.343 1202 4.0420.072 4.304 0.076 0.939 0.0149 580.5351 1203 1.531 0.200 2.192 0.1740.699 0.0166 789.5892 1202 1.755 0.049 1.952 0.065 0.899 0.0167 887.73521204 6.359 0.115 5.963 0.113 1.066 0.0174 828.5743 1202 5.301 0.0875.598 0.085 0.947 0.0178 916.7743 1204 5.952 0.113 5.571 0.107 1.0680.0178 856.672 1202 2.753 0.058 2.553 0.057 1.078 0.0179 827.5701 12026.362 0.095 6.684 0.092 0.952 0.0187 942.7879 1204 3.531 0.102 3.1950.092 1.105 0.019 523.4679 1203 3.471 0.153 4.011 0.165 0.865 0.0194915.7681 1204 6.365 0.115 5.972 0.116 1.066 0.0194 517.314 1101 5.5470.055 5.755 0.069 0.964 0.0195 777.553 1202 2.002 0.098 2.384 0.1300.840 0.0204 744.4956 1204 3.916 0.061 4.117 0.058 0.951 0.0206 440.35321204 1.502 0.136 0.975 0.180 1.539 0.021 782.5653 1201 6.382 0.065 6.6140.076 0.965 0.0222 494.4343 1203 2.145 0.226 2.900 0.232 0.740 0.0232303.1079 1202 5.646 0.048 5.802 0.047 0.973 0.0233 771.5814 1204 4.2080.050 4.367 0.046 0.964 0.0236 521.4522 1203 2.482 0.173 3.028 0.1580.820 0.0242 793.5681 1204 3.191 0.055 3.372 0.055 0.946 0.0244 832.60261102 3.662 0.042 3.795 0.040 0.965 0.0252 589.3403 1202 5.900 0.0846.171 0.085 0.956 0.026 732.4938 1204 4.319 0.061 4.511 0.057 0.9570.0263 783.5672 1101 3.993 0.085 4.259 0.079 0.938 0.0265 569.369 12024.947 0.042 5.089 0.047 0.972 0.0273 522.4635 1203 4.938 0.167 5.4850.176 0.900 0.0277 795.5181 1101 2.823 0.073 3.044 0.064 0.927 0.0287888.7394 1204 5.759 0.115 5.400 0.110 1.066 0.0287 490.3641 1203 1.8640.138 1.397 0.159 1.334 0.029 914.7583 1204 5.125 0.112 4.763 0.1181.076 0.0294 520.4499 1203 4.141 0.131 4.577 0.148 0.905 0.03 805.58321102 4.292 0.055 4.485 0.068 0.957 0.0301 795.5838 1204 3.558 0.0703.769 0.063 0.944 0.0305 575.2728 1101 2.532 0.060 2.739 0.074 0.9250.0324 831.5997 1102 4.774 0.042 4.903 0.040 0.974 0.0325 518.3174 11013.523 0.054 3.717 0.073 0.948 0.0338 520.3354 1101 2.271 0.117 2.6460.130 0.858 0.0354 573.4852 1203 4.919 0.063 4.740 0.053 1.038 0.0354579.5325 1203 4.235 0.135 4.636 0.127 0.913 0.0355 570.3725 1202 3.0210.043 3.155 0.045 0.958 0.0359 793.5386 1102 3.799 0.057 3.950 0.0390.962 0.0359 481.3172 1202 3.980 0.058 4.159 0.060 0.957 0.0361 831.57591101 4.485 0.066 4.677 0.060 0.959 0.0365 913.7513 1204 5.358 0.1135.006 0.120 1.070 0.037 772.5862 1204 3.306 0.045 3.437 0.042 0.9620.0378 304.241 1204 4.898 0.062 5.088 0.066 0.963 0.0397 746.557 12022.177 0.042 2.057 0.037 1.058 0.0399 519.3321 1101 4.014 0.116 4.3820.133 0.916 0.0405 833.7551 1204 2.611 0.191 3.140 0.161 0.831 0.0406889.7492 1204 7.711 0.122 7.365 0.110 1.047 0.0415 614.4914 1203 2.7470.091 2.448 0.114 1.122 0.0418 618.4829 1201 1.844 0.179 1.310 0.1851.407 0.0429 601.5164 1203 7.639 0.052 7.463 0.069 1.024 0.0431 671.57231204 2.349 0.088 2.604 0.087 0.902 0.0435 794.5421 1102 2.703 0.0592.853 0.040 0.948 0.0438 731.4916 1204 5.555 0.068 5.746 0.063 0.9670.0444 512.4082 1204 2.523 0.180 1.954 0.215 1.292 0.0451 768.5503 12022.028 0.082 2.277 0.090 0.891 0.0453 665.501 1204 3.429 0.105 3.0740.142 1.116 0.0454 890.7535 1204 7.055 0.116 6.731 0.106 1.048 0.0454759.5163 1204 4.804 0.055 4.954 0.048 0.970 0.0455 605.5457 1203 5.2350.100 5.509 0.089 0.950 0.0473 541.4425 1204 2.234 0.148 1.745 0.1981.280 0.0488 572.4816 1203 6.271 0.061 6.108 0.052 1.027 0.049 743.54661203 1.977 0.103 2.266 0.099 0.873 0.0492

TABLE 7 Accurate mass features differing between dementia patients witha significant cognitive impairment (ADAS ≧ 16) and dementia patientswith a mild cognitive impairment (ADAS ≦ 15) (p < 0.05, log2transformed). Detected Analysis AVG (log2) SEM AVG (log2) SEM log(2)Mass Mode ADAS high ADAS HIGH ADAS LOW ADAS LOW Ratio P Value 541.34321102 3.155 0.038 3.581 0.054 0.881 6.71E−09 567.3547 1102 2.633 0.0523.122 0.058 0.843 3.84E−08 569.3687 1102 2.079 0.053 2.526 0.054 0.8232.41E−07 829.5856 1102 3.861 0.057 4.303 0.059 0.897 1.71E−06 565.33941102 3.268 0.064 3.784 0.081 0.864 3.73E−06 837.5027 1101 3.373 0.0563.818 0.070 0.884 4.53E−06 804.5713 1102 4.022 0.051 4.414 0.060 0.9115.01E−06 831.5997 1102 4.379 0.055 4.761 0.046 0.920 5.17E−06 832.60261102 3.278 0.055 3.658 0.046 0.896 5.66E−06 803.568 1102 5.252 0.0525.640 0.065 0.931 1.42E−05 805.5832 1102 3.894 0.056 4.293 0.063 0.9071.48E−05 871.5528 1102 2.886 0.051 3.244 0.059 0.890 2.25E−05 555.31021102 1.669 0.053 2.070 0.074 0.806 2.34E−05 853.5854 1102 2.473 0.0582.824 0.040 0.876 2.85E−05 886.5582 1102 2.898 0.036 3.166 0.051 0.9164.34E−05 808.5792 1101 4.475 0.071 4.922 0.068 0.909 4.77E−05 780.54741101 5.229 0.067 5.665 0.086 0.923 0.0001 805.5605 1101 5.210 0.0725.655 0.078 0.921 0.0001 807.5758 1101 5.522 0.071 5.962 0.069 0.9260.0001 543.3296 1101 3.594 0.055 3.948 0.075 0.910 0.0002 779.5444 11016.228 0.067 6.660 0.086 0.935 0.0002 806.5639 1101 4.213 0.074 4.6560.079 0.905 0.0002 829.5604 1101 3.368 0.070 3.766 0.069 0.894 0.0002755.5468 1101 2.034 0.091 2.551 0.098 0.797 0.0003 819.5551 1102 2.2700.062 2.611 0.062 0.869 0.0003 915.5191 1101 2.209 0.066 2.603 0.0820.849 0.0004 794.5421 1102 2.389 0.054 2.705 0.073 0.883 0.0007 832.57911101 3.221 0.066 3.564 0.066 0.904 0.0007 793.5386 1102 3.462 0.0573.778 0.069 0.916 0.0008 803.5445 1101 4.995 0.065 5.351 0.078 0.9330.0008 804.5476 1101 4.045 0.064 4.402 0.080 0.919 0.0008 731.5464 11011.574 0.132 2.200 0.109 0.716 0.0011 827.57 1102 4.055 0.057 4.341 0.0620.934 0.0014 722.5335 1101 2.379 0.064 2.687 0.064 0.885 0.0015 831.57591101 4.163 0.072 4.510 0.070 0.923 0.0015 517.314 1101 5.343 0.057 5.6310.068 0.949 0.0019 757.4991 1101 3.393 0.086 3.828 0.102 0.886 0.0019783.5672 1101 3.547 0.087 3.974 0.097 0.893 0.002 545.3453 1101 3.4570.072 3.795 0.073 0.911 0.0022 518.3174 1101 3.321 0.056 3.600 0.0680.922 0.0023 760.5811 1101 4.181 0.092 4.609 0.101 0.907 0.003 784.58111101 4.155 0.104 4.630 0.111 0.897 0.0033 546.3485 1101 1.883 0.0672.196 0.078 0.857 0.0034 759.5779 1101 5.313 0.091 5.727 0.098 0.9280.0034 855.6016 1102 3.447 0.060 3.709 0.056 0.929 0.0034 847.5316 11012.627 0.075 3.004 0.106 0.874 0.0037 781.5617 1101 5.942 0.079 6.2910.082 0.945 0.0039 575.2728 1101 2.359 0.070 2.663 0.069 0.886 0.0042828.5737 1102 3.070 0.054 3.317 0.064 0.926 0.0047 795.555 1102 2.5580.065 2.831 0.063 0.904 0.0053 821.5712 1102 2.952 0.058 3.195 0.0570.924 0.0053 833.5932 1101 3.069 0.092 3.444 0.083 0.891 0.0053 782.5651101 4.918 0.082 5.266 0.085 0.934 0.0056 795.5181 1101 2.462 0.0752.793 0.090 0.882 0.0066 725.5527 1101 3.222 0.073 3.500 0.057 0.9210.0068 761.5843 1101 2.194 0.095 2.580 0.098 0.850 0.0078 738.5448 11022.493 0.076 2.766 0.047 0.901 0.0083 811.6096 1101 2.869 0.106 3.2770.096 0.876 0.0088 758.5656 1101 5.459 0.101 5.870 0.112 0.930 0.0092785.5933 1101 5.643 0.108 6.049 0.097 0.933 0.0103 458.2405 1101 1.6350.057 1.880 0.075 0.870 0.0104 757.5626 1101 6.418 0.100 6.815 0.1120.942 0.0112 541.3139 1101 3.968 0.074 4.278 0.096 0.928 0.0116 786.59671101 4.570 0.109 4.973 0.099 0.919 0.012 748.5721 1102 3.811 0.069 4.0500.055 0.941 0.0137 749.5761 1102 2.699 0.066 2.929 0.053 0.922 0.0156501.3212 1201 1.903 0.052 2.108 0.066 0.903 0.0176 809.5936 1101 4.7920.099 5.124 0.084 0.935 0.0194 810.5969 1101 3.785 0.102 4.126 0.0860.917 0.0199 542.3173 1101 2.183 0.077 2.464 0.091 0.886 0.0228 724.54931101 4.493 0.076 4.734 0.059 0.949 0.0232 723.5197 1204 3.966 0.0833.699 0.091 1.072 0.0362 521.3477 1101 3.545 0.086 3.863 0.130 0.9180.0374 1098.974 1204 3.016 0.105 2.552 0.219 1.182 0.039 523.3634 11013.263 0.100 3.596 0.130 0.907 0.0433 807.5768 1201 6.721 0.033 6.8160.031 0.986 0.0452 826.7069 1204 2.385 0.134 2.766 0.113 0.862 0.0462931.7695 1203 2.852 0.090 2.495 0.169 1.143 0.0475 183.0661 1101 2.3580.139 2.748 0.116 0.858 0.0485 504.3814 1203 1.818 0.065 1.573 0.1131.156 0.0489

TABLE 8 Accurate mass features differing between patients with mildcognitive impairment (MMSE 18-23), severe cognitive impairment (MMSE ≦17) and normal cognitive ability (MMSE ≧ 28) as measured on the MMSE.AVG SEM AVG SEM log (2) log(2) log(2) (log2) (log2) (log2) (log2) AVGSEM Ratio Ratio Ratio Detected Analysis mild mild severe severe (log2)(log2) mild/ severe/ mild/ Mass Mode MMSE MMSE MMSE MMSE Normal Normalnormal normal severe P Value 726.5461 1204 3.125 0.058 2.729 0.053 2.9810.067 1.048 0.916 1.145 0.0002 675.6377 1204 4.131 0.049 3.856 0.0824.124 0.061 1.002 0.935 1.071 0.0102 801.555* 1202 2.733 0.144 2.6020.110 3.052 0.077 0.895 0.853 1.050 0.0108 570.3725 1202 3.071 0.0672.895 0.054 3.130 0.054 0.981 0.925 1.061 0.0116 597.484 1203 2.9150.146 2.582 0.101 3.046 0.110 0.957 0.848 1.129 0.0138 246.1472 12042.437 0.187 3.031 0.161 2.974 0.067 0.819 1.019 0.804 0.0161 565.3394*1202 7.087 0.132 6.908 0.090 7.304 0.087 0.970 0.946 1.026 0.0171596.4791 1203 4.125 0.172 3.792 0.112 4.283 0.111 0.963 0.886 1.0880.0188 246.1465 1202 3.577 0.208 4.046 0.123 4.169 0.104 0.858 0.9700.884 0.0199 432.3254 1204 2.540 0.059 2.325 0.058 2.495 0.047 1.0180.932 1.092 0.0201 569.369* 1202 5.009 0.056 4.834 0.050 5.032 0.0630.995 0.961 1.036 0.0285 661.6233 1204 2.783 0.091 2.448 0.090 2.6830.079 1.037 0.912 1.137 0.0307 857.6186* 1202 3.923 0.122 3.699 0.1044.090 0.101 0.959 0.904 1.061 0.0332 804.5718 1202 5.940 0.135 5.7760.088 6.170 0.112 0.963 0.936 1.028 0.0334 803.5677 1202 7.274 0.1747.084 0.116 7.570 0.131 0.961 0.936 1.027 0.0359 542.3461 1202 3.8980.134 3.773 0.071 4.124 0.104 0.945 0.915 1.033 0.037 810.5399 12043.152 0.104 2.749 0.088 2.959 0.113 1.065 0.929 1.146 0.0374 728.56271204 3.233 0.078 2.954 0.065 3.054 0.071 1.059 0.967 1.094 0.0404566.3434 1202 5.338 0.100 5.142 0.093 5.460 0.082 0.978 0.942 1.0380.0417 481.3172 1202 3.940 0.088 3.875 0.069 4.136 0.077 0.953 0.9371.017 0.0436 724.5257 1204 3.168 0.102 2.875 0.079 2.961 0.055 1.0700.971 1.102 0.0447 825.5544 1202 2.933 0.173 2.605 0.144 3.160 0.1710.928 0.824 1.126 0.0474 826.5581 1202 2.005 0.151 1.579 0.156 2.1390.188 0.937 0.738 1.270 0.0488

TABLE 9 Grouping of patients into one of 8 groups based on the presenceof AD pathology, ADAS score and MMSE score. A score of 1 was given forthe presence of AD pathology, high ADAS score (≧16), or low MMSE score(≦23); a score of 0 was given in the absence of AD pathology, low ADASscore (≦15), or high MMSE score (≧28). AD Patient ID Pathology ADAS MMSE7001 1 1 1 7002 0 1 1 7006 1 0 1 7007 1 1 1 7008 1 0 1 7009 1 0 1 7010 10 1 7011 1 0 1 7014 1 0 1 7015 0 1 1 7016 1 1 1 7017 1 1 1 7022 1 1 17023 1 1 1 7024 1 0 1 7025 1 0 1 7027 1 0 1 7028 1 1 1 7029 1 1 1 7030 10 1 7031 1 0 1 7033 0 1 1 7034 1 0 1 7035 0 1 1 7037 1 1 1 7039 1 0 17042 1 0 1 7043 0 1 1 7044 1 1 1 7045 1 0 1 7046 1 1 1 7047 0 1 1 7048 10 1 7049 1 0 1 7050 0 1 1 7051 1 0 1 7052 1 0 1 7053 1 0 1 7055 0 1 17056 1 1 1 7057 1 0 1 7058 1 0 1 7059 0 1 1 7062 0 0 0 7063 1 1 1 7064 00 0 7066 0 0 1 7067 1 1 1 7070 0 0 0 7074 0 1 1 7075 1 1 1 7076 0 1 17077 1 1 1 7078 1 0 1 7079 0 1 1 7080 0 1 1 7081 1 1 1 7082 1 1 1 7084 11 1 7085 1 0 1 7086 0 1 1 7087 0 1 1 7088 1 1 1 7089 1 1 1 7090 1 1 17091 0 1 1 7093 1 0 1 7094 0 1 1 7095 0 1 1 7096 1 0 1 7097 1 0 1 7098 00 0 7101 0 0 0 7102 1 0 0 7103 0 1 0 7104 0 1 0 7105 0 1 0 7106 1 1 07108 0 0 0 7109 0 0 0 7110 0 1 0 7111 0 1 0 7112 1 1 0 7113 1 1 0 7114 11 0 7115 0 1 0 7116 1 1 0 7117 0 1 0 7118 0 0 0 7119 0 0 0 7120 0 0 07124 1 1 0 7125 1 1 0 7126 0 0 0 7127 0 0 0 7128 0 1 0 7129 1 0 0 7130 00 0 7131 0 0 0 7133 0 1 0 7134 0 1 0 7135 0 0 0 7136 1 0 0 7137 0 0 07138 0 0 0 7139 0 1 0 7140 1 0 0 7141 1 1 0 7142 1 1 0 7143 1 0 0 7144 10 0 7145 0 1 0 7146 0 1 0 7147 0 1 0 7150 1 0 0 7151 1 0 0 7152 0 1 07153 0 0 0 7154 1 1 0 7155 1 1 0 7156 0 0 0 7157 1 0 0 7158 1 0 0 7309 10 1 7310 1 0 0 7315 1 0 1 7802 0 0 0 7811 0 0 0 7814 1 0 0 7817 1 0 07818 0 0 0 7819 0 1 0 7823 1 0 0 7831 0 0 0 7832 1 0 0 7833 1 0 0

TABLE 10 Accurate mass features differing between patients showing thebest discrimination between AD and non-AD pathology (p < 0.05, log2transformed). AVG AVG Detected Analysis (log2) (log2) log(2) Mass ModeAD SEM AD non-AD SEM AD Ratio P Value 699.5199 1204 1.985 0.043 2.9560.067 0.672 1.24E−14 723.5195 1204 3.281 0.070 4.702 0.101 0.6984.86E−14 723.5194 1202 2.039 0.077 3.191 0.069 0.639 1.59E−13 751.55531204 4.290 0.074 5.450 0.093 0.787 5.86E−12 724.5258 1204 2.667 0.0623.750 0.092 0.711 6.50E−12 751.5529 1202 3.197 0.068 4.076 0.062 0.7841.03E−11 752.5564 1202 2.126 0.069 3.010 0.061 0.706 1.16E−11 749.53671202 3.061 0.062 3.861 0.056 0.793 1.22E−11 752.5581 1204 3.170 0.0724.292 0.094 0.739 1.52E−11 749.5406 1204 4.177 0.067 5.235 0.095 0.7984.77E−11 750.5403 1202 2.081 0.074 2.886 0.051 0.721 6.77E−11 750.54381204 3.060 0.068 4.067 0.091 0.752 7.99E−11 727.5567 1204 3.305 0.0744.064 0.053 0.813 4.71E−10 725.5385 1204 2.654 0.094 3.590 0.079 0.7393.99E−09 728.5629 1204 2.813 0.063 3.345 0.057 0.841 2.54E−07 726.54621204 2.753 0.064 3.215 0.062 0.856 7.09E−06 768.5538 1204 3.755 0.0604.250 0.084 0.884 2.34E−05 747.5244 1204 3.263 0.092 3.999 0.123 0.8162.45E−05 744.5535 1204 4.147 0.050 4.492 0.053 0.923 3.13E−05 813.58851202 2.969 0.051 3.362 0.067 0.883 3.58E−05 743.547 1204 5.096 0.0535.455 0.059 0.934 0.0001 767.5494 1204 4.835 0.065 5.337 0.092 0.9060.0001 775.5527 1204 2.993 0.100 3.645 0.105 0.821 0.0001 775.5533 12021.993 0.078 2.525 0.088 0.789 0.0001 811.5732 1202 2.590 0.072 3.0250.071 0.856 0.0001 812.5761 1202 1.599 0.064 2.057 0.084 0.777 0.0001817.5376 1102 1.992 0.073 2.490 0.081 0.800 0.0001 747.5201 1202 1.8290.090 2.412 0.110 0.758 0.0002 795.5837 1204 3.418 0.073 3.810 0.0620.897 0.0002 796.5874 1204 2.468 0.051 2.839 0.074 0.869 0.0002 748.52861204 1.812 0.247 2.917 0.126 0.621 0.0003 810.54 1204 2.627 0.088 3.1160.083 0.843 0.0003 782.5084 1204 3.308 0.060 3.687 0.078 0.897 0.0004755.486 1204 3.069 0.080 3.477 0.073 0.883 0.0005 758.5092 1204 4.4180.065 4.707 0.043 0.939 0.0007 771.5813 1204 3.965 0.074 4.288 0.0460.925 0.0007 304.241 1204 4.716 0.057 5.013 0.062 0.941 0.0011 814.59191202 1.988 0.046 2.307 0.077 0.862 0.0011 786.5415 1204 3.671 0.0743.979 0.050 0.923 0.0013 787.5728 1202 1.743 0.085 2.136 0.075 0.8160.0013 793.5386 1102 3.337 0.086 3.690 0.057 0.904 0.0015 783.5147 12043.166 0.071 3.512 0.077 0.901 0.0022 828.5736 1102 2.906 0.080 3.2920.087 0.883 0.0023 828.5743 1202 5.132 0.073 5.485 0.081 0.936 0.0024743.5468 1203 1.812 0.118 2.235 0.061 0.811 0.0029 784.5235 1204 3.1960.081 3.490 0.045 0.916 0.003 760.5215 1204 3.910 0.072 4.200 0.0580.931 0.0032 787.5464 1204 2.617 0.090 2.937 0.050 0.891 0.0037 827.571102 3.933 0.078 4.313 0.096 0.912 0.0039 305.2438 1204 2.384 0.0642.648 0.058 0.900 0.0041 794.5421 1102 2.275 0.090 2.601 0.058 0.8750.0041 827.5701 1202 6.193 0.081 6.559 0.090 0.944 0.0046 590.343 12023.939 0.055 4.216 0.074 0.934 0.0047 589.3404 1202 5.769 0.061 6.0760.085 0.950 0.0057 789.5893 1202 1.673 0.064 1.967 0.077 0.851 0.0057670.5689 1204 3.048 0.095 3.510 0.127 0.868 0.006 855.6016 1102 3.3250.079 3.646 0.079 0.912 0.0064 759.5162 1204 4.510 0.086 4.811 0.0600.937 0.0065 671.5722 1204 2.157 0.123 2.635 0.111 0.819 0.0066 769.56541204 3.829 0.065 4.086 0.063 0.937 0.0069 856.6063 1202 4.446 0.0764.777 0.087 0.931 0.007 409.0208 1202 2.638 0.057 2.899 0.073 0.9100.0072 774.0313 1204 2.317 0.202 1.370 0.268 1.691 0.0076 804.5713 11023.964 0.086 4.276 0.071 0.927 0.008 855.6025 1202 5.437 0.074 5.7490.085 0.946 0.0083 530.382 1204 2.991 0.089 2.097 0.309 1.426 0.0085626.5278 1203 3.676 0.068 3.915 0.053 0.939 0.0087 731.4914 1204 5.2810.086 5.589 0.073 0.945 0.0093 732.4937 1204 4.050 0.083 4.348 0.0700.931 0.0093 517.3722 1204 2.189 0.112 1.465 0.241 1.494 0.0096 741.53071204 2.754 0.086 3.117 0.105 0.884 0.0109 803.5681 1102 5.187 0.0875.491 0.073 0.945 0.0111 746.5714 1204 2.962 0.065 3.228 0.076 0.9180.0113 627.5304 1203 2.544 0.068 2.783 0.058 0.914 0.0118 544.397 12043.249 0.120 2.323 0.341 1.399 0.0145 745.5656 1204 3.812 0.071 4.0910.083 0.932 0.0148 739.5142 1204 2.576 0.105 3.009 0.137 0.856 0.0162570.3725 1202 2.879 0.050 3.071 0.057 0.938 0.0164 686.4877 1204 2.5270.068 2.757 0.064 0.917 0.0179 517.3136 1201 2.907 0.104 2.575 0.0881.129 0.0193 744.4956 1204 3.741 0.073 3.988 0.070 0.938 0.0202 529.31671202 2.929 0.057 3.199 0.095 0.916 0.0203 821.5713 1102 2.825 0.0893.091 0.065 0.914 0.0207 819.5641 1202 2.632 0.117 3.063 0.136 0.8590.0209 667.5474 1204 3.336 0.118 2.882 0.150 1.158 0.0222 886.5582 11022.820 0.064 3.038 0.067 0.928 0.0232 796.529 1204 2.815 0.070 3.0630.078 0.919 0.0234 306.2569 1204 2.848 0.083 3.103 0.074 0.918 0.0256581.3344 1202 1.676 0.132 2.082 0.115 0.805 0.0261 817.584 1202 2.1810.064 2.383 0.060 0.915 0.0261 685.5538 1204 1.862 0.132 2.222 0.0830.838 0.0263 555.3101 1102 1.640 0.094 1.944 0.093 0.844 0.0277 666.54551204 4.533 0.128 4.059 0.168 1.117 0.0301 820.5677 1202 1.550 0.1632.037 0.145 0.761 0.031 773.5368 1202 1.455 0.109 1.808 0.114 0.8050.0324 541.3432 1102 3.148 0.073 3.377 0.073 0.932 0.0328 767.547 12022.917 0.095 3.237 0.111 0.901 0.0345 569.369 1202 4.824 0.048 4.9870.057 0.967 0.0352 871.5935 1202 1.702 0.068 2.049 0.145 0.830 0.03621226.0968 1203 3.089 0.116 2.709 0.130 1.140 0.0365 793.5679 1204 3.0220.067 3.237 0.075 0.934 0.0407 411.3211 1202 2.817 0.068 2.988 0.0420.943 0.0408 684.5491 1204 2.304 0.087 2.600 0.110 0.886 0.041 768.55041202 1.786 0.141 2.160 0.106 0.827 0.041 743.5469 1202 2.894 0.076 3.1730.109 0.912 0.0417 482.3215 1202 1.866 0.054 2.056 0.072 0.908 0.0424574.4637 1202 1.059 0.201 1.653 0.204 0.640 0.0444 509.3493 1202 2.2900.057 2.490 0.079 0.919 0.0446 772.586 1204 3.167 0.062 3.330 0.0500.951 0.0458 383.3283 1204 1.280 0.142 1.662 0.119 0.770 0.0462 664.53221204 4.643 0.145 4.176 0.173 1.112 0.0465 312.2312 1204 2.771 0.0592.583 0.070 1.073 0.0467 481.3172 1202 3.803 0.057 3.982 0.065 0.9550.0468 765.5335 1204 3.065 0.107 3.418 0.137 0.897 0.0491 847.5953 12022.162 0.114 2.518 0.134 0.858 0.0492 624.5131 1203 3.900 0.068 4.0840.061 0.955 0.0497

TABLE 11 Accurate mass features differing between patients showing thebest discrimination between high ADAS score and low ADAS score (p <0.05, log2 transformed). AVG AVG Detected Analysis (log2) SEM high(log2) SEM low log(2) Mass Mode high ADAS ADAS low ADAS ADAS Ratio PValue 541.3435 1102 3.007 0.047 3.741 0.083 0.804 5.76E−09 569.3685 11021.862 0.065 2.686 0.070 0.693 9.27E−09 804.5713 1102 3.915 0.047 4.4940.080 0.871 3.59E−07 803.5681 1102 5.145 0.046 5.720 0.084 0.8995.31E−07 837.5027 1101 3.257 0.058 3.866 0.070 0.842 5.33E−07 807.57581101 5.318 0.076 6.045 0.065 0.880 5.92E−07 808.5792 1101 4.265 0.0764.992 0.069 0.854 7.70E−07 832.6024 1102 3.129 0.065 3.740 0.063 0.8371.34E−06 779.5444 1101 6.061 0.061 6.691 0.082 0.906 1.39E−06 780.54741101 5.073 0.063 5.714 0.083 0.888 1.42E−06 831.5995 1102 4.245 0.0684.850 0.061 0.875 2.37E−06 546.3486 1101 1.593 0.073 2.250 0.084 0.7085.48E−06 871.5529 1102 2.778 0.050 3.307 0.090 0.840 6.59E−06 567.35451102 2.637 0.079 3.305 0.081 0.798 8.54E−06 545.3453 1101 3.142 0.0853.836 0.077 0.819 9.04E−06 805.5604 1101 5.013 0.074 5.685 0.099 0.8829.51E−06 794.5422 1102 2.287 0.056 2.795 0.079 0.818 1.22E−05 886.55841102 2.853 0.053 3.354 0.081 0.851 1.32E−05 827.57 1102 3.893 0.0634.438 0.079 0.877 1.41E−05 828.5737 1102 2.920 0.061 3.425 0.069 0.8521.56E−05 805.5831 1102 3.821 0.058 4.385 0.100 0.871 1.61E−05 829.58561102 3.781 0.076 4.393 0.077 0.861 1.68E−05 915.5191 1101 2.144 0.0532.621 0.078 0.818 1.99E−05 793.5387 1102 3.352 0.056 3.854 0.086 0.8702.76E−05 806.5637 1101 4.001 0.078 4.667 0.107 0.857 2.85E−05 821.57131102 2.834 0.067 3.309 0.046 0.856 3.30E−05 565.3396 1102 3.216 0.0963.992 0.121 0.806 3.57E−05 555.3098 1102 1.608 0.081 2.242 0.094 0.7174.04E−05 757.4989 1101 3.119 0.086 3.849 0.148 0.810 0.0001 832.57921101 3.024 0.092 3.662 0.085 0.826 0.0001 855.6015 1102 3.274 0.0683.795 0.093 0.862 0.0001 458.2404 1101 1.430 0.076 1.984 0.109 0.7210.0002 783.5671 1101 3.275 0.105 4.025 0.141 0.813 0.0002 803.5445 11014.778 0.095 5.397 0.079 0.885 0.0002 804.5475 1101 3.844 0.095 4.4590.075 0.862 0.0002 831.576 1101 3.924 0.100 4.575 0.095 0.858 0.0002518.3174 1101 3.093 0.072 3.609 0.101 0.857 0.0003 781.5616 1101 5.6790.096 6.315 0.110 0.899 0.0003 517.314 1101 5.122 0.077 5.642 0.1040.908 0.0004 782.5649 1101 4.649 0.100 5.296 0.118 0.878 0.0004 785.59331101 5.333 0.121 6.107 0.132 0.873 0.0004 786.5967 1101 4.256 0.1245.039 0.133 0.844 0.0004 833.5932 1101 2.746 0.113 3.494 0.134 0.7860.0004 853.5855 1102 2.333 0.088 2.875 0.080 0.811 0.0004 755.5468 11011.879 0.101 2.593 0.157 0.725 0.0005 811.6096 1101 2.512 0.140 3.3750.152 0.744 0.0006 809.5936 1101 4.448 0.126 5.177 0.119 0.859 0.0007761.5843 1101 1.993 0.094 2.654 0.160 0.751 0.0008 810.5969 1101 3.4510.128 4.183 0.115 0.825 0.0008 847.5316 1101 2.543 0.093 3.082 0.0920.825 0.0008 757.5626 1101 6.144 0.104 6.825 0.155 0.900 0.0009 758.56561101 5.185 0.107 5.885 0.158 0.881 0.0009 543.3296 1101 3.486 0.0844.025 0.125 0.866 0.001 759.5779 1101 5.080 0.100 5.759 0.169 0.8820.0011 760.5811 1101 3.952 0.102 4.645 0.177 0.851 0.0012 795.5551 11022.493 0.075 2.937 0.092 0.849 0.0012 829.5604 1101 3.200 0.098 3.7590.106 0.851 0.0012 523.3635 1101 2.882 0.117 3.595 0.163 0.801 0.0014575.2728 1101 2.204 0.078 2.673 0.104 0.824 0.0014 784.581 1101 3.9210.129 4.667 0.171 0.840 0.0018 819.5553 1102 2.210 0.089 2.665 0.0850.829 0.0025 817.5378 1102 2.089 0.088 2.491 0.068 0.839 0.0047 731.54631101 1.373 0.174 2.218 0.197 0.619 0.0052 795.5183 1101 2.270 0.0812.725 0.140 0.833 0.0058 722.5334 1101 2.190 0.085 2.598 0.111 0.8430.0083 760.5217 1204 4.084 0.063 4.331 0.042 0.943 0.0118 833.757 12032.531 0.161 3.274 0.231 0.773 0.0132 429.3749 1204 2.703 0.080 3.1370.167 0.861 0.0143 722.4789 1201 1.895 0.093 2.247 0.064 0.843 0.01451228.1111 1203 2.699 0.127 3.291 0.194 0.820 0.0146 541.3139 1101 3.9090.116 4.412 0.152 0.886 0.0158 860.7753 1203 4.152 0.110 4.647 0.1610.893 0.016 703.5709 1101 2.858 0.095 3.216 0.072 0.888 0.0163 579.53251203 3.776 0.177 4.492 0.191 0.841 0.0166 858.686 1202 3.030 0.064 2.7810.057 1.090 0.0166 831.7408 1203 3.708 0.137 4.286 0.176 0.865 0.0175484.3041 1203 2.257 0.120 1.475 0.348 1.530 0.0176 580.5352 1203 1.3100.222 2.135 0.172 0.614 0.0181 830.7362 1203 4.568 0.137 5.156 0.1870.886 0.0182 495.332 1101 4.769 0.114 5.305 0.195 0.899 0.0186 183.06621101 1.868 0.215 2.671 0.183 0.699 0.0191 702.5677 1101 4.033 0.1044.409 0.074 0.915 0.0194 887.8001 1203 2.884 0.137 3.396 0.122 0.8490.0199 886.7916 1203 4.153 0.131 4.631 0.120 0.897 0.0232 759.5164 12044.697 0.077 4.975 0.066 0.944 0.0236 725.5527 1101 3.123 0.108 3.4990.079 0.892 0.0244 832.7522 1203 3.418 0.170 4.119 0.245 0.830 0.0255828.7207 1203 5.336 0.130 5.867 0.187 0.909 0.0263 496.3355 1101 2.9720.113 3.473 0.195 0.856 0.0264 604.5432 1203 6.338 0.149 6.863 0.1370.924 0.0283 864.738 1204 4.641 0.166 5.295 0.232 0.876 0.0298 680.56231204 3.590 0.125 4.126 0.214 0.870 0.0305 627.5201 1204 3.756 0.1574.380 0.227 0.858 0.0315 1019.3838 1102 3.116 0.042 2.950 0.063 1.0560.0323 825.6927 1203 1.408 0.212 2.138 0.204 0.658 0.0331 863.7336 12045.382 0.170 6.035 0.237 0.892 0.0338 521.3477 1101 3.368 0.126 3.8900.213 0.866 0.0349 731.4916 1204 5.513 0.065 5.746 0.076 0.959 0.0351542.3173 1101 2.116 0.120 2.565 0.157 0.825 0.0352 829.7242 1204 2.5750.161 3.185 0.221 0.809 0.0356 835.6998 1204 2.285 0.247 3.153 0.2710.725 0.0356 276.2095 1204 2.479 0.091 2.885 0.179 0.859 0.0358 653.5361204 4.096 0.154 4.679 0.218 0.875 0.0368 523.4678 1203 2.747 0.2383.609 0.295 0.761 0.0372 829.7246 1203 4.606 0.121 5.065 0.173 0.9090.0376 921.8142 1204 2.618 0.338 3.654 0.184 0.716 0.039 605.5456 12034.920 0.154 5.420 0.131 0.908 0.0392 549.4844 1203 5.315 0.129 5.7530.137 0.924 0.0398 732.4939 1204 4.258 0.065 4.480 0.069 0.950 0.04365.3159 1203 1.907 0.098 1.449 0.217 1.316 0.0402 867.7581 1204 2.3580.280 3.299 0.284 0.715 0.0402 738.5445 1102 2.458 0.122 2.825 0.0590.870 0.0409 859.7695 1203 5.769 0.102 6.185 0.179 0.933 0.0414 625.50751204 3.271 0.138 3.811 0.224 0.858 0.0422 512.3356 1203 2.602 0.0952.151 0.223 1.210 0.0432 842.7387 1203 2.626 0.103 2.998 0.139 0.8760.0432 552.5021 1203 2.742 0.161 3.315 0.213 0.827 0.0444 724.5493 11014.410 0.105 4.736 0.081 0.931 0.0447 626.5108 1204 2.117 0.159 2.6770.209 0.791 0.0454 928.7505 1203 2.869 0.168 2.238 0.265 1.282 0.0473894.781 1204 3.884 0.196 4.542 0.228 0.855 0.048 866.7533 1204 3.9050.219 4.667 0.292 0.837 0.0492 278.2255 1204 4.693 0.070 5.006 0.1560.937 0.0494 865.7486 1204 4.644 0.223 5.419 0.296 0.857 0.0497

TABLE 12 Accurate mass features differing between patients showing thebest discrimination between high MMSE score and low MMSE score (p <0.05, log2 transformed). AVG AVG (log2) SEM (log2) SEM Detected Analysislow low high high log(2) Mass Mode MMSE MMSE MMSE MMSE Ratio P Value505.3229 1202 3.585 0.045 4.514 0.053 0.794 3.90E−15 857.6186 1202 3.3390.058 4.298 0.053 0.777 5.05E−14 803.5678 1202 6.747 0.056 7.962 0.0820.847 5.19E−14 858.6211 1202 2.300 0.058 3.221 0.059 0.714 6.73E−13566.3434 1202 4.856 0.055 5.682 0.055 0.855 1.90E−12 801.555 1202 2.2200.057 3.128 0.064 0.710 2.89E−12 832.6036 1202 4.692 0.057 5.569 0.0610.842 3.64E−12 804.5718 1202 5.516 0.047 6.475 0.080 0.852 4.63E−12565.3393 1202 6.629 0.054 7.462 0.062 0.888 9.30E−12 506.3214 1202 2.2370.051 2.908 0.044 0.769 1.26E−11 743.5467 1202 2.635 0.077 3.621 0.0660.728 2.57E−11 831.6 1202 5.793 0.055 6.774 0.087 0.855 4.69E−11 744.551202 1.608 0.082 2.504 0.056 0.642 1.72E−10 570.3724 1202 2.702 0.0423.289 0.052 0.821 4.09E−10 829.5859 1202 5.572 0.065 6.476 0.085 0.8608.42E−10 830.5894 1202 4.477 0.061 5.311 0.078 0.843 1.01E−09 509.34941202 2.113 0.042 2.685 0.059 0.787 4.17E−09 542.3461 1202 3.562 0.0474.292 0.082 0.830 6.29E−09 711.2577 1202 1.891 0.072 2.585 0.057 0.7311.12E−08 709.2595 1202 1.864 0.079 2.546 0.047 0.732 1.49E−08 481.31721202 3.642 0.055 4.233 0.060 0.860 2.74E−08 569.369 1202 4.664 0.0465.203 0.063 0.897 7.82E−08 805.5839 1202 5.216 0.051 5.867 0.082 0.8891.16E−07 806.5873 1202 4.024 0.052 4.658 0.078 0.864 1.16E−07 856.60611202 4.179 0.069 5.099 0.121 0.819 1.81E−07 743.5469 1204 5.166 0.0515.635 0.049 0.917 1.96E−07 793.5386 1102 2.966 0.065 3.468 0.043 0.8552.63E−07 855.6023 1202 5.166 0.065 6.090 0.128 0.848 2.81E−07 482.32151202 1.742 0.057 2.291 0.066 0.761 4.15E−07 541.3433 1202 5.318 0.0676.197 0.123 0.858 4.56E−07 744.5535 1204 4.203 0.048 4.631 0.048 0.9084.68E−07 807.5899 1202 2.124 0.071 2.730 0.070 0.778 8.81E−07 827.57011202 5.937 0.077 6.931 0.147 0.857 1.20E−06 847.5954 1202 2.030 0.0952.867 0.104 0.708 1.37E−06 828.5742 1202 4.908 0.074 5.818 0.137 0.8441.67E−06 787.5464 1204 2.601 0.059 3.069 0.056 0.847 2.17E−06 591.35421202 3.749 0.081 4.387 0.078 0.855 3.05E−06 794.5422 1102 1.910 0.0652.338 0.040 0.817 3.76E−06 804.5713 1102 3.619 0.065 4.092 0.056 0.8854.94E−06 803.568 1102 4.829 0.068 5.317 0.057 0.908 5.22E−06 758.50941204 4.437 0.052 4.813 0.046 0.922 5.89E−06 592.3571 1202 2.013 0.0842.634 0.079 0.764 7.14E−06 760.5216 1204 3.930 0.056 4.313 0.045 0.9118.02E−06 759.5165 1204 4.558 0.067 5.001 0.050 0.911 9.01E−06 767.54691202 2.700 0.119 3.526 0.101 0.766 9.49E−06 786.5416 1204 3.664 0.0514.093 0.064 0.895 1.12E−05 784.5239 1204 3.102 0.069 3.575 0.060 0.8681.23E−05 821.5711 1102 2.415 0.067 2.863 0.055 0.843 1.25E−05 819.56411202 2.451 0.109 3.287 0.126 0.746 2.03E−05 777.5531 1202 1.473 0.1332.257 0.083 0.653 2.21E−05 853.5862 1202 4.524 0.069 5.222 0.126 0.8663.29E−05 507.3316 1202 2.653 0.056 3.061 0.064 0.867 3.70E−05 731.49171204 5.356 0.062 5.744 0.053 0.932 4.18E−05 831.5998 1102 3.958 0.0814.429 0.057 0.894 4.26E−05 854.5902 1202 3.521 0.075 4.231 0.130 0.8324.70E−05 630.5586 1203 2.845 0.054 3.242 0.069 0.877 0.0001 732.494 12044.127 0.062 4.492 0.050 0.919 0.0001 741.5307 1204 2.661 0.099 3.3370.105 0.797 0.0001 771.5815 1204 4.036 0.064 4.423 0.055 0.913 0.0001819.5552 1102 1.733 0.081 2.149 0.045 0.806 0.0001 832.6027 1102 2.8640.081 3.318 0.056 0.863 0.0001 871.5527 1102 2.467 0.073 2.914 0.0660.846 0.0001 871.5935 1202 1.550 0.108 2.224 0.110 0.697 0.0001 886.79171203 3.826 0.095 4.459 0.099 0.858 0.0001 910.7968 1203 3.257 0.0833.757 0.079 0.867 0.0001 529.3166 1202 2.771 0.071 3.332 0.112 0.8320.0002 589.3404 1202 5.578 0.072 6.318 0.155 0.883 0.0002 768.5503 12021.668 0.144 2.412 0.103 0.691 0.0002 884.7801 1203 6.076 0.082 6.6390.104 0.915 0.0002 568.3574 1202 3.760 0.055 4.144 0.074 0.907 0.0003685.2601 1202 1.483 0.102 2.052 0.093 0.723 0.0003 765.5313 1202 1.4470.148 2.219 0.120 0.652 0.0003 772.5863 1204 3.177 0.051 3.466 0.0500.917 0.0003 829.5856 1102 3.407 0.088 3.862 0.064 0.882 0.0003 911.80321203 2.388 0.077 2.794 0.064 0.855 0.0003 786.5967 1101 4.302 0.0964.955 0.132 0.868 0.0004 885.7854 1203 5.116 0.080 5.615 0.095 0.9110.0004 174.1409 1203 1.904 0.122 2.872 0.212 0.663 0.0005 590.3431 12023.790 0.071 4.415 0.142 0.858 0.0005 604.5432 1203 6.172 0.114 6.8030.113 0.907 0.0005 757.5626 1101 6.170 0.099 6.804 0.129 0.907 0.0005879.5999 1202 2.601 0.104 3.235 0.124 0.804 0.0005 605.5456 1203 4.7620.115 5.393 0.118 0.883 0.0006 757.5016 1204 3.549 0.094 3.977 0.0570.893 0.0006 581.3345 1202 1.572 0.124 2.253 0.131 0.698 0.0007 628.54211203 3.231 0.070 3.606 0.070 0.896 0.0007 723.5195 1202 2.196 0.0852.730 0.110 0.805 0.0007 749.5365 1202 3.146 0.084 3.579 0.078 0.8790.0007 785.5933 1101 5.382 0.098 6.025 0.137 0.893 0.0007 887.7999 12032.493 0.171 3.227 0.087 0.773 0.0007 886.8012 1204 3.209 0.117 3.7820.096 0.849 0.0008 186.1408 1203 1.539 0.100 2.279 0.172 0.675 0.0009200.1564 1203 2.724 0.075 3.280 0.129 0.830 0.0009 769.5655 1204 3.8330.075 4.197 0.063 0.913 0.0009 817.5838 1202 2.041 0.086 2.452 0.0700.833 0.0009 715.5169 1204 2.238 0.200 3.073 0.108 0.728 0.001 699.521204 2.223 0.082 2.690 0.099 0.826 0.0011 758.5656 1101 5.231 0.1015.844 0.134 0.895 0.0011 795.5182 1101 2.324 0.087 2.800 0.098 0.8300.0011 772.528 1204 3.202 0.075 3.531 0.053 0.907 0.0012 739.5164 12021.109 0.170 1.859 0.124 0.597 0.0014 779.5444 1101 6.186 0.098 6.6390.083 0.932 0.0014 244.2187 1203 2.645 0.062 3.037 0.093 0.871 0.0015508.3356 1202 0.969 0.088 1.386 0.078 0.700 0.0015 811.6096 1101 2.5690.101 3.204 0.152 0.802 0.0017 882.766 1203 7.238 0.092 7.755 0.1170.933 0.0017 744.4954 1204 3.832 0.067 4.118 0.049 0.930 0.0018 884.78771204 4.658 0.101 5.127 0.092 0.909 0.0018 262.2293 1203 2.639 0.0592.992 0.084 0.882 0.002 784.5811 1101 3.896 0.100 4.493 0.143 0.8670.002 817.5375 1102 1.715 0.076 2.148 0.101 0.798 0.002 723.5197 12043.643 0.087 4.211 0.143 0.865 0.0021 810.5401 1204 2.693 0.070 3.1190.104 0.863 0.0021 880.6035 1202 1.837 0.084 2.351 0.126 0.782 0.0021807.5757 1101 5.434 0.097 5.912 0.102 0.919 0.0022 883.7705 1203 6.3960.085 6.870 0.111 0.931 0.0022 749.576 1102 2.516 0.057 2.174 0.0841.157 0.0023 852.5738 1202 3.365 0.120 3.968 0.135 0.848 0.0025 752.55641202 2.277 0.101 2.730 0.092 0.834 0.0027 783.5672 1101 3.273 0.1063.856 0.141 0.849 0.0027 808.5792 1101 4.393 0.099 4.868 0.104 0.9020.0027 755.5468 1101 1.885 0.107 2.412 0.121 0.782 0.0031 780.5475 11015.202 0.096 5.629 0.089 0.924 0.0031 826.707 1204 2.266 0.118 2.7430.086 0.826 0.0031 851.5698 1202 4.297 0.118 4.898 0.142 0.877 0.0031631.5619 1203 1.475 0.132 1.956 0.066 0.754 0.0033 755.4861 1204 3.0190.074 3.424 0.102 0.882 0.0034 188.1566 1203 2.272 0.134 2.926 0.1540.777 0.0035 883.7765 1204 4.148 0.099 4.589 0.095 0.904 0.0035 726.54591204 2.790 0.064 3.090 0.069 0.903 0.0038 750.5403 1202 2.185 0.0892.549 0.073 0.857 0.0038 627.5302 1203 2.497 0.080 2.845 0.075 0.8780.0039 885.7922 1204 3.954 0.103 4.382 0.089 0.902 0.0039 675.6357 12031.815 0.210 0.815 0.240 2.228 0.0042 751.5529 1202 3.374 0.098 3.7870.090 0.891 0.0043 882.7723 1204 4.984 0.102 5.422 0.096 0.919 0.0043530.3213 1202 1.209 0.089 1.698 0.129 0.712 0.0044 380.3096 1204 1.4320.107 1.867 0.092 0.767 0.0046 783.5148 1204 3.121 0.064 3.517 0.1110.887 0.0048 202.172 1203 3.048 0.112 3.631 0.153 0.839 0.0049 748.57211102 3.632 0.058 3.325 0.082 1.092 0.0049 603.5317 1203 7.585 0.0777.883 0.060 0.962 0.005 243.0718 1101 4.430 0.144 3.210 0.373 1.3800.0052 582.2473 1201 3.377 0.132 2.673 0.191 1.264 0.0055 739.5141 12042.498 0.195 3.173 0.109 0.787 0.0056 879.7454 1204 3.291 0.124 3.7490.088 0.878 0.0057 494.4342 1203 1.382 0.353 2.777 0.303 0.497 0.0059782.5086 1204 3.266 0.073 3.662 0.110 0.892 0.0059 855.6016 1102 2.9840.080 3.363 0.097 0.887 0.006 921.8153 1204 2.175 0.312 3.187 0.1310.682 0.006 724.5256 1204 2.877 0.077 3.324 0.128 0.866 0.0061 183.06611101 2.155 0.151 2.748 0.130 0.784 0.0065 522.4634 1203 4.325 0.2385.237 0.198 0.826 0.0067 853.7296 1204 3.607 0.109 4.012 0.083 0.8990.0068 216.1877 1203 2.796 0.092 3.282 0.140 0.852 0.0076 779.5442 12017.005 0.079 7.301 0.065 0.959 0.0076 777.5288 1201 2.657 0.093 3.0090.079 0.883 0.0077 523.468 1203 2.895 0.227 3.731 0.183 0.776 0.0083569.3687 1102 1.734 0.098 2.075 0.067 0.836 0.0085 775.5535 1202 2.1120.083 2.438 0.079 0.866 0.0087 632.5035 1204 4.858 0.122 5.325 0.1100.912 0.0088 805.5604 1101 5.112 0.099 5.504 0.097 0.929 0.0089 313.2691203 1.592 0.175 2.180 0.112 0.730 0.009 743.5464 1203 1.703 0.150 2.1560.056 0.790 0.009 768.5539 1204 3.881 0.077 4.240 0.100 0.915 0.009686.488 1204 2.671 0.054 2.872 0.047 0.930 0.0092 886.5582 1102 2.5030.058 2.781 0.079 0.900 0.0092 602.5282 1203 8.833 0.081 9.116 0.0590.969 0.0093 558.4667 1202 1.932 0.201 2.652 0.159 0.728 0.0095 809.59361101 4.523 0.112 5.054 0.155 0.895 0.0101 881.7616 1204 4.015 0.1114.427 0.099 0.907 0.0101 833.7558 1204 1.919 0.311 2.863 0.138 0.6700.0103 864.7596 1203 2.184 0.067 2.468 0.078 0.885 0.0103 613.3405 12023.422 0.124 3.963 0.150 0.863 0.0104 626.5277 1203 3.649 0.073 3.9460.079 0.925 0.0105 629.5449 1203 2.015 0.088 2.370 0.093 0.850 0.0106767.5494 1204 4.975 0.083 5.343 0.104 0.931 0.0106 517.3722 1204 2.1760.149 1.408 0.235 1.545 0.0107 773.5347 1204 3.151 0.056 3.426 0.0820.920 0.0108 806.5638 1101 4.113 0.100 4.498 0.097 0.914 0.0109 204.18761203 2.205 0.099 2.682 0.143 0.822 0.0113 837.5027 1101 3.439 0.0753.734 0.078 0.921 0.0116 595.4932 1202 1.393 0.217 2.170 0.185 0.6420.0117 915.5192 1101 2.301 0.087 2.588 0.060 0.889 0.0117 257.8105 11013.636 0.084 2.677 0.344 1.358 0.0121 541.343 1102 2.816 0.087 3.1050.062 0.907 0.0121 745.5656 1204 3.838 0.051 4.054 0.062 0.947 0.0125749.5408 1204 4.465 0.071 4.865 0.130 0.918 0.0126 725.5385 1204 2.9320.068 3.293 0.115 0.890 0.0128 757.4993 1101 3.262 0.095 3.716 0.1400.878 0.0128 852.725 1204 3.774 0.105 4.122 0.076 0.916 0.0128 780.54741201 5.941 0.081 6.223 0.067 0.955 0.0129 880.7516 1203 7.205 0.1077.651 0.127 0.942 0.0129 881.7558 1203 6.471 0.103 6.883 0.116 0.9400.0132 338.2821 1203 4.474 0.095 4.792 0.073 0.934 0.0135 827.5699 11023.597 0.089 3.987 0.116 0.902 0.0136 880.7566 1204 4.483 0.117 4.8820.094 0.918 0.0138 572.4468 1204 1.726 0.131 2.213 0.129 0.780 0.014336.2662 1203 3.181 0.097 3.518 0.083 0.904 0.0141 615.3539 1202 2.2630.089 2.598 0.092 0.871 0.0146 544.4479 1203 1.928 0.085 2.273 0.1000.848 0.0148 579.5325 1203 3.737 0.170 4.362 0.166 0.857 0.0148 520.451203 3.613 0.209 4.378 0.204 0.825 0.0149 633.5078 1204 3.891 0.1284.347 0.118 0.895 0.0149 519.3321 1101 3.416 0.099 3.875 0.146 0.8810.0151 547.6031 1101 2.454 0.114 1.714 0.259 1.432 0.0152 727.5569 12043.548 0.077 3.866 0.093 0.918 0.0152 339.285 1203 2.242 0.107 2.6010.087 0.862 0.0154 746.5719 1204 3.004 0.054 3.196 0.050 0.940 0.0154828.5736 1102 2.605 0.083 2.966 0.111 0.878 0.0154 798.6742 1203 1.6850.154 2.329 0.192 0.723 0.0155 860.7753 1203 4.054 0.129 4.507 0.1170.899 0.0156 521.4522 1203 1.837 0.271 2.786 0.244 0.660 0.0158 594.48781202 2.699 0.253 3.523 0.191 0.766 0.0158 781.5617 1101 5.737 0.1106.187 0.134 0.927 0.0158 810.5969 1101 3.498 0.121 4.023 0.161 0.8700.0158 631.4902 1204 2.690 0.150 3.208 0.132 0.839 0.0163 518.4346 12031.198 0.238 2.121 0.267 0.565 0.0164 244.0559 1101 3.875 0.099 3.3590.174 1.153 0.0165 751.5556 1204 4.690 0.092 5.091 0.126 0.921 0.0165825.5544 1202 2.647 0.146 3.202 0.159 0.827 0.017 520.3353 1101 1.6610.103 2.114 0.144 0.786 0.0176 858.7632 1203 6.769 0.125 7.244 0.1370.935 0.0178 861.7801 1203 2.123 0.171 2.681 0.138 0.792 0.0179 646.48371204 1.688 0.192 2.245 0.108 0.752 0.0184 489.6451 1101 2.555 0.1091.822 0.269 1.402 0.0186 255.8135 1101 4.518 0.090 3.502 0.393 1.2900.019 260.2135 1203 1.422 0.129 1.878 0.128 0.757 0.0193 833.5931 11012.952 0.114 3.347 0.109 0.882 0.0197 831.5759 1101 4.038 0.114 4.4510.121 0.907 0.0201 544.3971 1204 3.313 0.158 2.377 0.342 1.394 0.0203294.2207 1204 5.248 0.117 5.652 0.112 0.929 0.0205 724.5493 1101 4.6740.050 4.395 0.101 1.063 0.0205 663.487 1204 1.733 0.223 2.369 0.1280.731 0.0207 860.7753 1204 3.460 0.170 3.955 0.104 0.875 0.0207 246.14651202 3.707 0.141 4.188 0.134 0.885 0.0209 606.4869 1204 4.562 0.1375.023 0.126 0.908 0.0215 576.5097 1203 9.304 0.133 9.756 0.127 0.9540.0216 311.775 1101 2.594 0.152 1.849 0.264 1.403 0.022 566.413 12042.026 0.190 2.553 0.100 0.794 0.0221 577.5136 1203 7.959 0.125 8.3920.124 0.948 0.0224 835.7001 1204 2.246 0.249 3.007 0.187 0.747 0.0226665.501 1204 2.974 0.116 3.370 0.114 0.883 0.0227 675.6377 1204 3.8250.077 4.086 0.075 0.936 0.023 600.5127 1203 8.597 0.082 8.866 0.0750.970 0.0232 752.5583 1204 3.555 0.096 3.939 0.126 0.902 0.0241 161.10531101 4.011 0.148 2.977 0.402 1.347 0.0242 616.4673 1201 1.329 0.2182.031 0.193 0.654 0.0245 253.8163 1101 4.184 0.090 3.283 0.364 1.2740.0248 878.74 1204 3.665 0.118 4.001 0.074 0.916 0.025 580.535 12031.221 0.219 1.936 0.203 0.631 0.0252 859.7694 1203 5.668 0.122 6.0760.119 0.933 0.0252 538.4224 1202 1.087 0.172 1.689 0.184 0.643 0.0256804.723 1203 1.425 0.264 2.238 0.214 0.637 0.0256 833.7571 1203 2.5990.192 3.201 0.165 0.812 0.0261 530.382 1204 2.988 0.204 2.114 0.3051.413 0.0262 908.7832 1203 5.011 0.090 5.309 0.088 0.944 0.027 653.53611204 3.973 0.129 4.367 0.107 0.910 0.0273 559.47 1202 0.812 0.174 1.3880.171 0.585 0.0274 748.5735 1202 3.935 0.071 3.726 0.054 1.056 0.0279803.5445 1101 4.899 0.121 5.305 0.124 0.924 0.028 832.7521 1203 3.4340.205 4.059 0.169 0.846 0.0283 320.2357 1204 1.225 0.148 1.609 0.0710.761 0.0284 728.5626 1204 2.941 0.066 3.161 0.067 0.930 0.0284 795.5551102 2.053 0.080 2.292 0.062 0.896 0.0285 838.7232 1204 2.011 0.2532.753 0.191 0.730 0.0285 782.565 1101 4.709 0.114 5.143 0.147 0.9160.0286 750.544 1204 3.351 0.066 3.678 0.124 0.911 0.0287 783.4315 11012.505 0.125 1.807 0.271 1.386 0.029 214.172 1203 3.549 0.090 3.910 0.1260.908 0.0292 312.2663 1203 3.879 0.140 4.315 0.126 0.899 0.03 857.75311203 7.332 0.109 7.738 0.138 0.947 0.03 202.0453 1101 5.911 0.122 5.2380.266 1.128 0.0305 763.5157 1202 1.570 0.175 2.098 0.147 0.748 0.0306218.0192 1101 4.023 0.134 3.143 0.357 1.280 0.0307 856.7481 1203 8.1730.118 8.608 0.151 0.949 0.032 795.5839 1204 3.545 0.061 3.800 0.0930.933 0.0321 565.4104 1204 3.353 0.123 3.754 0.127 0.893 0.0332 313.77221101 3.080 0.178 2.250 0.319 1.369 0.0334 909.7882 1203 4.177 0.0924.456 0.081 0.938 0.0337 429.6888 1101 2.475 0.093 1.833 0.268 1.3510.0341 431.957 1202 1.847 0.154 2.312 0.136 0.799 0.0341 607.4919 12042.906 0.259 3.579 0.147 0.812 0.0341 294.1443 1201 2.449 0.210 1.7950.201 1.364 0.0348 598.4965 1203 6.590 0.104 6.923 0.106 0.952 0.035549.6005 1101 2.027 0.167 1.424 0.212 1.424 0.0351 283.9028 1101 3.0180.076 2.397 0.267 1.259 0.0353 820.5679 1202 1.438 0.153 2.061 0.2310.698 0.0353 574.4597 1204 3.354 0.210 3.994 0.196 0.840 0.0359 826.70481203 4.509 0.145 4.950 0.134 0.911 0.0361 805.5832 1102 3.502 0.0853.742 0.066 0.936 0.0364 550.4956 1203 6.603 0.188 7.165 0.167 0.9220.0365 630.4861 1204 3.807 0.132 4.238 0.142 0.898 0.0369 551.4985 12035.182 0.182 5.733 0.167 0.904 0.0371 549.4845 1203 5.363 0.140 5.7650.115 0.930 0.0372 827.7083 1203 3.751 0.145 4.174 0.124 0.899 0.0374306.2569 1204 2.821 0.075 3.051 0.072 0.925 0.0375 775.553 1204 3.2720.071 3.577 0.119 0.915 0.0377 242.203 1203 4.260 0.087 4.573 0.1130.932 0.0382 828.7202 1204 3.465 0.158 3.906 0.122 0.887 0.0385 232.21881203 3.156 0.098 3.499 0.122 0.902 0.0387 722.4789 1201 1.873 0.0842.132 0.082 0.879 0.0389 530.3471 1202 2.566 0.117 2.910 0.103 0.8820.0393 264.2452 1203 1.753 0.138 2.114 0.090 0.829 0.0397 858.7663 12045.791 0.140 6.169 0.100 0.939 0.0401 759.5779 1101 5.093 0.108 5.4710.136 0.931 0.0404 559.469 1204 2.761 0.191 3.288 0.149 0.840 0.0406593.4743 1204 2.069 0.240 2.728 0.184 0.758 0.0406 392.2938 1204 1.9410.277 2.718 0.225 0.714 0.041 575.499 1203 7.133 0.096 7.452 0.112 0.9570.0414 826.5581 1202 1.759 0.125 2.203 0.163 0.798 0.0417 719.6233 12044.578 0.094 4.279 0.102 1.070 0.0419 295.2287 1204 2.883 0.203 3.4140.138 0.844 0.0422 555.3101 1102 1.310 0.070 1.541 0.080 0.850 0.0424830.7355 1204 3.947 0.183 4.447 0.141 0.888 0.0427 474.3706 1203 0.8680.128 1.234 0.112 0.704 0.0433 203.1157 1101 3.694 0.116 2.965 0.3191.246 0.0437 295.2239 1204 3.109 0.123 3.476 0.119 0.894 0.0437 855.74171204 4.834 0.122 5.184 0.107 0.932 0.0438 760.5811 1101 3.973 0.1094.349 0.138 0.913 0.0441 625.5076 1204 3.218 0.140 3.624 0.126 0.8880.0443 270.2343 1203 1.714 0.106 2.018 0.095 0.850 0.0444 661.6233 12042.375 0.073 2.644 0.103 0.898 0.0445 446.3403 1202 0.751 0.166 1.2750.181 0.589 0.0446 521.3474 1201 2.493 0.169 2.031 0.137 1.228 0.0451837.718 1204 2.369 0.329 3.272 0.267 0.724 0.0451 228.1877 1203 2.9540.108 3.313 0.131 0.892 0.0458 832.7495 1204 2.994 0.226 3.588 0.1640.834 0.0458 576.477 1202 1.734 0.213 2.337 0.187 0.742 0.0459 1085.32941101 3.756 0.022 3.463 0.136 1.085 0.0467 444.2717 1202 1.195 0.0961.496 0.105 0.799 0.0473 545.6062 1101 1.955 0.134 1.427 0.213 1.3700.0477 651.5221 1204 3.677 0.131 4.039 0.111 0.910 0.0482 829.7244 12034.651 0.135 5.044 0.130 0.922 0.0482 859.7706 1204 4.962 0.145 5.3310.100 0.931 0.0482 575.4632 1204 2.109 0.207 2.692 0.185 0.783 0.0485804.5476 1101 3.984 0.119 4.353 0.130 0.915 0.0485 725.5527 1101 3.4070.058 3.160 0.102 1.078 0.0486 350.2423 1202 2.544 0.227 1.901 0.2071.338 0.0489 382.1083 1101 2.600 0.210 1.858 0.286 1.399 0.049 836.70761204 1.427 0.242 2.163 0.256 0.660 0.0493 671.5726 1204 2.055 0.1382.458 0.136 0.836 0.0495 197.8549 1101 3.431 0.102 2.697 0.337 1.2720.0499 773.537 1202 1.569 0.078 1.805 0.082 0.869 0.0499

TABLE 13 Accurate mass features differing between clinically diagnosedAD and non-AD patients in CSF (p < 0.05, log2 transformed). AVG AVGDetected Analysis (log2) SEM (log2) SEM log(2) Mass Mode AD AD non-ADnon-AD Ratio P Value 742.2972 1203 1.187 0.130 0.000 0.000 1.1871.69E−05 562.46 1203 1.080 0.134 0.000 0.000 1.080 4.84E−05 731.653 12030.905 0.193 0.000 0.000 0.905 0.0022 432.1532 1203 1.445 0.125 0.8800.053 0.609 0.0037 487.6482 1101 2.388 0.044 2.668 0.057 1.117 0.0037275.8712 1101 2.362 0.057 2.664 0.058 1.128 0.0045 371.7311 1101 3.4170.098 3.818 0.022 1.117 0.0053 622.2539 1203 2.667 0.110 2.166 0.0740.812 0.0056 485.6503 1101 1.758 0.051 1.976 0.024 1.124 0.0058 207.08221203 1.010 0.126 0.198 0.198 0.196 0.006 640.2637 1203 3.557 0.152 2.8760.106 0.809 0.0063 373.728 1101 3.013 0.086 3.358 0.037 1.114 0.0071656.2587 1203 0.977 0.080 0.322 0.199 0.330 0.0097 730.6493 1203 1.8880.139 0.948 0.278 0.502 0.0109 220.0798 1101 2.252 0.078 2.556 0.0441.135 0.011 641.2661 1203 1.915 0.148 1.010 0.282 0.527 0.0152 779.43931101 3.612 0.032 3.728 0.022 1.032 0.0167 782.5647 1201 1.898 0.2081.128 0.142 0.594 0.0172 313.7721 1101 4.045 0.096 4.350 0.031 1.0750.0215 785.4288 1101 3.273 0.039 3.410 0.033 1.042 0.0253 777.4426 11012.495 0.017 2.614 0.044 1.048 0.0279 341.8614 1101 2.747 0.069 2.9900.061 1.089 0.0305 250.0366 1101 3.122 0.115 3.462 0.049 1.109 0.0309315.7693 1101 3.347 0.090 3.618 0.044 1.081 0.0311 253.8164 1101 4.2370.108 4.542 0.030 1.072 0.033 369.7338 1101 2.713 0.148 3.132 0.0351.154 0.034 781.5607 1201 2.728 0.234 2.048 0.095 0.751 0.0345 206.07891203 4.178 0.147 3.484 0.253 0.834 0.0357 429.689 1101 3.232 0.042 3.3920.051 1.050 0.0358 638.2465 1203 1.927 0.188 1.372 0.095 0.712 0.0362578.1574 1101 1.828 0.042 1.674 0.046 0.916 0.0368 262.0777 1101 4.7380.114 5.072 0.052 1.070 0.0371 265.8423 1101 3.682 0.101 3.960 0.0281.076 0.0395 582.6334 1101 1.702 0.051 1.526 0.052 0.897 0.0395 255.81351101 4.678 0.113 4.978 0.029 1.064 0.0418 321.8039 1101 3.190 0.0983.480 0.062 1.091 0.0423 514.6879 1101 2.650 0.037 2.534 0.032 0.9560.0424 262.0221 1101 3.633 0.126 3.964 0.026 1.091 0.0435 746.621 11011.015 0.225 0.306 0.190 0.301 0.0438 781.4354 1101 4.163 0.028 4.2540.025 1.022 0.045 311.775 1101 3.457 0.111 3.764 0.053 1.089 0.0461257.8105 1101 3.753 0.115 4.056 0.036 1.081 0.0472

TABLE 14 Retention Time of Six Biomarkers Q-Star Retention FT AccurateTheoretical Mass Time Neutral Mass Formula Neutral Mass (M − H) ion(min) 541.3432 C25H52NO9P 541.3379 540.3616 29.4 569.3687 C27H56NO9P569.3692 568.3979 31.0 699.5198 C39H74NO7P 699.5202 698.5392 40.5723.5195 C41H74NO7P 723.5202 722.5331 40.2 751.5555 C43H78NO7P 751.5515750.5667 41.9 803.568 C43H82NO10P 803.5676 802.5575 38.6

TABLE 15 Metabolite 541.3432 Fragments Frag Q-Star Formula TheoreticalDetected Delta Diff Loss C25H51NO9P 540.33014 540.3616 0.0315 — —C23H47NO7P 480.30901 480.3313 0.0223 60.02112 C2H4O2 C16H31O2 255.23240255.2521 0.0197 292.15251 C9H27NO7P C7H15NO5P 224.06878 224.0904 0.0216300.26644 C18H36O3 O3P 78.95850 78.975 0.0165 461.37163 C25H51NO6Legend: frag formula: The putative computationally derived molecularformula of the fragment neutral mass. theoretical: The theoretical massof the formulas shown in the frag formula column. Qstar-detected: Thedetected mass from the ABI Q-Star XL. delta: The difference between thetheoretical and neutral mass. diff: The mass difference between theQstar-detected parent ion mass and the Qstar-detected fragmant ion mass.Loss: The putative molecular formulas of the “diff” column Note: Theseare only predicted formulas for each fragment and are not necessarilythe actual formulas.

TABLE 16 Metabolite 569.3687 Fragments Q-Star Frag Formula TheoreticalDetected Delta Diff Loss C27H55NO9P 568.3614 568.3979 0.0365 — —C25H51NO7P 508.34031 508.3612 0.0209 60.02112 C2H4O2 C18H35O2 283.26370283.2900 0.0263 285.09773 C9H20NO7P C7H15NO5P 224.06878 224.0899 0.0211344.23266 C20H40O4 C4H11NO4P 168.04256 168.0668 0.0242 400.31887C23H44O5 C3H6PO5 152.99528 153.0196 0.0243 446.33992 C24H49NO4P O3P78.95850 78.975 0.0165 489.40293 C27H55NO6 Legend: frag formula: Theputative computationally derived molecular formula of the fragmentneutral mass. theoretical: The theoretical mass of the formulas shown inthe frag formula column. Qstar-detected: The detected mass from the ABIQ-Star XL. delta: The difference between the theoretical and neutralmass. diff: The mass difference between the Qstar-detected parent ionmass and the Qstar-detected fragmant ion mass. Loss: The putativemolecular formulas of the “diff” column Note: These are only predictedformulas for each fragment and are not necessarily the actual formulas.

TABLE 17 Metabolite 803.568 Fragments Q-Star Frag Formula TheoreticalDetected Delta Differences Loss C43H81NO10P 802.55980 802.5575 −0.0023 —— C41H77NO8P 742.53868 742.5526 0.0139 76.01604 C2H4O3 C23H47NO7P480.30901 480.3248 0.0158 322.25079 C20H34O3 C18H32O2 279.23240 279.24850.0161 522.31957 C25H49NO8P C16H31O2 255.23240 255.2469 0.0145 547.32740C27H50NO8P C7H15NO5P 224.06878 224.0853 0.0165 578.49102 C36H66O5C4H11NO4P 168.04256 168.0629 0.0203 634.51724 C39H70O6 Legend: fragformula: The putative computationally derived molecular formula of thefragment neutral mass. theoretical: The theoretical mass of the formulasshown in the frag formula column. Qstar-detected: The detected mass fromthe ABI Q-Star XL. delta: The difference between the theoretical andneutral mass. diff: The mass difference between the Qstar-detectedparent ion mass and the Qstar-detected fragmant ion mass. Loss: Theputative molecular formulas of the “diff” column Note: These are onlypredicted formulas for each fragment and are not necessarily the actualformula

TABLE 18 List of Preferred List of Metabolites Metabolite CodeMetabolite Name Molecular Formula Parent Mass M − H Mass DiagnosticFragment Mass MS/MS Transition M01 PtdEt 16:0/18:0 C39H78N1O8P1719.54648 718.5 R1 (C16H31O2) - 255 718.0/255.0 M02 PtdEt 16:0/18:1C39H76N1O8P1 717.53083 716.5 R1 (C16H31O2) - 255 716.0/255.0 M03 PtdEt18:0/18:0 C41H82N1O8P1 747.57777 746.5 R1 (C18H35O2) - 283 746.0/283.0M04 PtdEt 18:0/18:1 C41H80N1O8P1 745.56213 744.5 R1 (C18H35O2) - 283744.0/283.0 M05 Plasmanyl 16:0/18:1 C39H78N1O7P1 703.55156 702.5 R2(C18H33O2) - 281 702.0/281.0 M06 Plasmanyl 16:0/18:2 C39H76N1O7P1701.53591 700.5 R2 (C18H31O2) - 279 700.0/279.0 M07 Plasmanyl 16:0/20:4C41H76N1O7P1 725.53591 724.5 R2 (C20H31O2) - 303 724.0/303.0 M08Plasmanyl 16:0/22:4 C43H80N1O7P1 753.56721 752.5 R2 (C22H35O2) - 331752.0/331.0 M09 Plasmanyl 16:0/22:6 C43H76N1O7P1 749.53591 748.5 R2(C22H31O2) - 327 748.0/327.0 M10 Plasmanyl 18:0/18:1 C41H82N1O7P1731.58286 730.5 R2 (C18H33O2) - 281 730.0/281.0 M11 Plasmanyl 18:0/18:2C41H80N1O7P1 729.56721 728.5 R2 (C18H31O2) - 279 728.0/279.0 M12Plasmanyl 18:0/20:4 C43H80N1O7P1 753.56721 752.5 R2 (C20H31O2) - 303752.0/303.0 M13 Plasmanyl 18:0/22:4 C45H84N1O7P1 781.59851 780.5 R2(C22H35O2) - 331 780.0/331.0 M14 Plasmanyl 18:0/22:6 C45H80N1O7P1777.56721 776.5 R2 (C22H31O2) - 327 776.0/327.0 M15 Plasmenyl 16:0/18:1C39H76N1O7P1 701.53591 700.5 R2 (C18H33O2) - 281 700.0/281.0 M16Plasmenyl 16:0/18:2 C39H74N1O7P1 699.52026 698.5 R2 (C18H31O2) - 279698.0/279.0 M17 Plasmenyl 16:0/20:4 C41H74N1O7P1 723.52026 722.5 R2(C20H31O2) - 303 722.0/303.0 M18 Plasmenyl 16:0/22:4 C43H78N1O7P1751.55156 750.5 R2 (C22H35O2) - 331 750.0/331.0 M19 Plasmenyl 16:0/22:6C43H74N1O7P1 747.52026 746.5 R2 (C22H31O2) - 327 746.0/327.0 M20Plasmenyl 18:0/18:1 C41H80N1O7P1 729.56721 728.5 R2 (C18H33O2) - 281728.0/281.0 M21 Plasmenyl 18:0/18:2 C41H78N1O7P1 727.55156 726.5 R2(C18H31O2) - 279 726.0/279.0 M22 Plasmenyl 18:0/20:4 C43H78N1O7P1751.55156 750.5 R2 (C20H31O2) - 303 750.6/303.2 M23 Plasmenyl 18:0/22:4C45H82N1O7P1 779.58286 778.5 R2 (C22H35O2) - 331 778.0/331.0 M24Plasmenyl 18:0/22:6 C45H78N1O7P1 775.55156 774.5 R2 (C22H31O2) - 327774.0/327.0 M25 Free 22:6 C22H32O2 328.24022 327.2 (C21H31) - 283327.2/283.0 M26 Free 20:4 C20H32O2 304.24022 303.2 (C19H31) - 259303.2/259.5

TABLE 19 Clinical Data on Subject Cohorts Age MMSE ADAS-cog Population nMean SEM Mean SEM Mean SEM Age Ctl, 30-39, Female 14 36.4 0.9 Age Ctl,30-39, Male 11 35.2 1.0 Age Ctl, 40-49, Female 44 44.8 0.5 Age Ctl,40-49, Male 27 44.7 0.6 Age Ctl, 50-59, Female 107 54.2 0.3 Age Ctl,50-59, Male 59 54.1 0.4 Age Ctl, 60-69, Female 55 63.4 0.3 Age Ctl,60-69, Male 34 64.4 0.5 Age Ctl, 70+_Female 27 79.7 1.2 Age Ctl,70+_Male 35 75.5 0.7 Cognitive Normal, Female 36 77.6 1.1 29.6 0.1Cognitive Normal, Male 32 76.8 1.1 29.3 0.1 SDAT_all, Female 140 80.00.6 12.6 0.7 34.2 1.6 SDAT_all, Male 117 79.8 0.7 15.3 0.5 27.4 1.3SDAT, ADAS 5-19, Female 38 79.6 1.2 17.6 0.7 15.2 0.6 SDAT, ADAS 20-39,Female 54 78.6 1.0 16.6 0.7 27.0 0.8 SDAT, ADAS 40-70, Female 48 81.91.1 4.2 0.7 57.3 1.5 SDAT, ADAS 5-19, Male 40 79.0 1.1 17.3 0.7 15.3 0.5SDAT, ADAS 20-39, Male 58 79.6 0.9 16.8 0.6 27.5 0.7 SDAT, ADAS 40-70,Male 18 82.6 2.1 6.2 1.1 53.2 2.2 Post Mortem SDAT Male 10 80.1 1.4 PostMortem SDAT Female 10 77.6 1.5 Post Mortem Ctl, Female 9 84.4 1.8 PostMortem Ctl, Male 10 77.9 1.4

TABLE 20 Effect of Age on Serum Ethanolamine Phospholipid Levels inMales Age Ctl, 30-39, Male Age Ctl, 40-49, Male Age Ctl, 50-59, Male AgeCtl, 60-69, Male Age Ctl, 70+_Male Metabolite Code Mean SEM Mean SEMMean SEM Mean SEM Mean SEM M01 0.122 0.017 0.119 0.008 0.113 0.006 0.1320.007 0.130 0.006 M02 0.056 0.008 0.058 0.006 0.058 0.004 0.059 0.0070.056 0.005 M03 0.102 0.014 0.085 0.006 0.095 0.005 0.103 0.008 0.1100.010 M04 0.026 0.004 0.027 0.003 0.025 0.002 0.027 0.003 0.027 0.003M05 0.014 0.002 0.012 0.001 0.011 0.001 0.012 0.001 0.011 0.001 M060.032 0.005 0.026 0.002 0.026 0.001 0.025 0.002 0.027 0.002 M07 0.0780.015 0.053 0.007 0.063 0.005 0.061 0.007 0.060 0.006 M08 0.010 0.0020.007 0.001 0.007 0.001 0.007 0.001 0.006 0.001 M09 0.019 0.004 0.0130.002 0.019 0.002 0.019 0.002 0.022 0.003 M10 0.058 0.009 0.052 0.0040.049 0.003 0.055 0.004 0.053 0.003 M11 0.131 0.021 0.099 0.007 0.0980.005 0.099 0.007 0.104 0.007 M12 0.244 0.037 0.186 0.015 0.210 0.0120.213 0.017 0.202 0.016 M13 0.015 0.002 0.013 0.001 0.013 0.001 0.0130.001 0.012 0.001 M14 0.034 0.005 0.026 0.003 0.036 0.002 0.040 0.0040.047 0.006 M15 0.085 0.012 0.075 0.005 0.074 0.004 0.074 0.006 0.0760.005 M16 0.291 0.043 0.220 0.016 0.244 0.013 0.223 0.018 0.246 0.018M17 0.489 0.102 0.332 0.035 0.429 0.029 0.412 0.048 0.410 0.044 M180.032 0.005 0.025 0.003 0.027 0.002 0.026 0.004 0.023 0.002 M19 0.0860.014 0.063 0.008 0.094 0.008 0.098 0.012 0.114 0.017 M20 0.086 0.0130.070 0.006 0.068 0.004 0.069 0.006 0.071 0.006 M21 0.294 0.044 0.2360.017 0.247 0.014 0.222 0.018 0.236 0.018 M22 1.054 0.220 0.788 0.0790.930 0.067 0.933 0.102 0.906 0.104 M23 0.022 0.003 0.019 0.001 0.0190.001 0.020 0.003 0.017 0.001 M24 0.077 0.014 0.063 0.008 0.086 0.0070.088 0.010 0.103 0.016 M25 0.205 0.014 0.195 0.017 0.274 0.018 0.3100.025 0.374 0.031 M26 0.091 0.011 0.082 0.005 0.096 0.003 0.109 0.0090.114 0.009

TABLE 21 Ratio and T-test values between males of different ages 50-59vs. 40-49, Male 60-69 vs. 40-49, Male 70+ vs. 40-49, Male MetaboliteCode Ratio ttest Ratio ttest Ratio ttest M01 0.955 6.0E−01 1.110 2.2E−011.097 2.4E−01 M02 0.998 9.9E−01 1.024 8.8E−01 0.965 8.1E−01 M03 1.1252.3E−01 1.213 9.4E−02 1.296 5.0E−02 M04 0.954 7.0E−01 1.019 9.1E−011.015 9.3E−01 M05 0.953 6.3E−01 1.011 9.3E−01 0.979 8.5E−01 M06 1.0317.5E−01 0.976 8.3E−01 1.054 6.2E−01 M07 1.198 2.4E−01 1.164 4.1E−011.133 4.8E−01 M08 1.031 8.2E−01 1.014 9.4E−01 0.907 5.4E−01 M09 1.4432.7E−02 1.473 4.5E−02 1.701 2.1E−02 M10 0.953 5.9E−01 1.061 5.9E−011.013 8.9E−01 M11 0.992 9.3E−01 1.004 9.7E−01 1.058 5.6E−01 M12 1.1282.5E−01 1.147 2.5E−01 1.085 4.8E−01 M13 1.007 9.4E−01 1.032 8.0E−010.921 4.4E−01 M14 1.383 1.5E−02 1.536 7.0E−03 1.802 2.8E−03 M15 0.9878.9E−01 0.985 8.9E−01 1.020 8.5E−01 M16 1.109 2.7E−01 1.013 9.1E−011.118 2.9E−01 M17 1.291 5.4E−02 1.241 2.1E−01 1.235 1.9E−01 M18 1.0556.6E−01 1.032 8.7E−01 0.904 4.9E−01 M19 1.484 1.9E−02 1.540 2.6E−021.789 1.9E−02 M20 0.971 7.8E−01 0.976 8.4E−01 1.014 9.0E−01 M21 1.0486.3E−01 0.941 5.8E−01 1.000 1.0E+00 M22 1.179 2.1E−01 1.183 2.9E−011.149 3.9E−01 M23 1.008 9.3E−01 1.066 7.5E−01 0.900 3.6E−01 M24 1.3655.2E−02 1.403 6.0E−02 1.642 4.7E−02 M25 1.405 9.0E−03 1.589 6.7E−041.912 1.7E−05 M26 1.175 1.8E−02 1.324 1.7E−02 1.389 6.3E−03

TABLE 22 Effect of Age on Serum Ethanolamine Phospholipid Levels inFemales Age Ctl, 30-39, Female Age Ctl, 40-49, Female Age Ctl, 50-59,Female Age Ctl, 60-69, Female Age Ctl, 70+_Female Metabolite Code MeanSEM Mean SEM Mean SEM Mean SEM Mean SEM M01 0.103 0.012 0.114 0.0060.105 0.006 0.127 0.007 0.126 0.009 M02 0.047 0.006 0.069 0.006 0.0580.004 0.058 0.004 0.056 0.004 M03 0.095 0.015 0.101 0.007 0.093 0.0050.091 0.006 0.100 0.005 M04 0.028 0.005 0.033 0.003 0.025 0.002 0.0230.002 0.026 0.002 M05 0.011 0.002 0.013 0.001 0.010 0.000 0.010 0.0010.011 0.001 M06 0.031 0.005 0.031 0.002 0.025 0.001 0.023 0.001 0.0250.002 M07 0.061 0.010 0.071 0.007 0.057 0.003 0.051 0.005 0.052 0.005M08 0.008 0.001 0.010 0.001 0.007 0.000 0.006 0.001 0.006 0.001 M090.021 0.004 0.021 0.002 0.020 0.001 0.017 0.002 0.019 0.002 M10 0.0500.007 0.059 0.003 0.049 0.002 0.052 0.003 0.051 0.003 M11 0.120 0.0180.119 0.008 0.100 0.004 0.099 0.006 0.100 0.008 M12 0.196 0.022 0.2220.015 0.191 0.009 0.181 0.011 0.188 0.013 M13 0.013 0.001 0.015 0.0010.012 0.001 0.011 0.001 0.012 0.001 M14 0.040 0.007 0.043 0.004 0.0420.003 0.041 0.003 0.049 0.004 M15 0.071 0.009 0.079 0.005 0.068 0.0030.066 0.004 0.069 0.004 M16 0.274 0.044 0.268 0.021 0.226 0.011 0.2100.013 0.232 0.017 M17 0.419 0.076 0.458 0.053 0.392 0.024 0.354 0.0360.360 0.033 M18 0.028 0.004 0.032 0.003 0.025 0.002 0.019 0.002 0.0230.003 M19 0.107 0.024 0.100 0.012 0.099 0.007 0.087 0.009 0.103 0.008M20 0.074 0.013 0.075 0.006 0.063 0.003 0.063 0.004 0.068 0.005 M210.294 0.057 0.275 0.024 0.228 0.012 0.210 0.014 0.229 0.020 M22 0.9380.170 1.025 0.108 0.853 0.055 0.759 0.068 0.837 0.096 M23 0.021 0.0030.022 0.002 0.017 0.001 0.015 0.001 0.017 0.002 M24 0.102 0.025 0.0930.011 0.089 0.006 0.081 0.008 0.096 0.008 M25 0.239 0.023 0.236 0.0140.277 0.012 0.326 0.022 0.373 0.020 M26 0.091 0.012 0.093 0.006 0.0930.003 0.094 0.004 0.107 0.007

TABLE 23 Ratio and T-test values between females of different ages 50-59vs. 40-49, Female 60-69 vs. 40-49, Female 70+ vs. 40-49, FemaleMetabolite Code Ratio ttest Ratio ttest Ratio ttest M01 0.925 3.7E−011.118 1.8E−01 1.110 2.6E−01 M02 0.838 1.3E−01 0.835 9.5E−02 0.8121.1E−01 M03 0.927 4.0E−01 0.906 2.9E−01 0.989 9.0E−01 M04 0.772 2.1E−020.709 5.1E−03 0.811 1.2E−01 M05 0.811 1.1E−02 0.798 2.0E−02 0.8328.7E−02 M06 0.815 1.1E−02 0.731 1.5E−03 0.805 5.7E−02 M07 0.798 3.4E−020.718 1.9E−02 0.726 4.9E−02 M08 0.721 4.2E−03 0.569 1.3E−04 0.6401.9E−02 M09 0.934 5.8E−01 0.784 9.4E−02 0.912 5.6E−01 M10 0.841 3.1E−020.882 1.3E−01 0.863 1.1E−01 M11 0.843 2.3E−02 0.832 3.5E−02 0.8431.1E−01 M12 0.862 6.2E−02 0.815 2.5E−02 0.848 1.2E−01 M13 0.806 8.8E−030.737 2.0E−03 0.839 1.3E−01 M14 0.981 8.6E−01 0.953 6.9E−01 1.1492.6E−01 M15 0.864 5.6E−02 0.841 3.7E−02 0.871 1.5E−01 M16 0.846 5.3E−020.786 1.5E−02 0.867 2.3E−01 M17 0.856 2.0E−01 0.773 1.0E−01 0.7861.9E−01 M18 0.778 2.4E−02 0.594 1.8E−04 0.733 7.4E−02 M19 0.986 9.2E−010.872 3.8E−01 1.030 8.5E−01 M20 0.843 5.6E−02 0.842 9.5E−02 0.9054.0E−01 M21 0.829 5.0E−02 0.763 1.8E−02 0.832 1.9E−01 M22 0.832 1.2E−010.741 3.3E−02 0.817 2.3E−01 M23 0.797 1.5E−02 0.663 3.3E−04 0.7767.4E−02 M24 0.957 7.4E−01 0.871 3.6E−01 1.040 8.1E−01 M25 1.176 4.8E−021.381 1.7E−03 1.581 2.4E−07 M26 1.003 9.6E−01 1.021 7.9E−01 1.1591.2E−01

TABLE 24 Effect of Dementia State on Serum Ethanolamine PhospholipidLevels in Females Cognitive Normal, SDAT_all, SDAT, ADAS SDAT, ADASSDAT, ADAS Female Female 5-19, Female 20-39, Female 40-70, FemaleMetabolite Code Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM M01 0.1180.005 0.113 0.003 0.109 0.004 0.112 0.005 0.118 0.006 M02 0.062 0.0050.053 0.002 0.050 0.003 0.057 0.004 0.052 0.004 M03 0.099 0.005 0.0760.002 0.080 0.004 0.076 0.004 0.073 0.004 M04 0.026 0.002 0.025 0.0010.024 0.002 0.027 0.002 0.024 0.003 M05 0.011 0.001 0.009 0.000 0.0100.001 0.009 0.001 0.009 0.001 M06 0.027 0.001 0.022 0.001 0.024 0.0010.023 0.001 0.020 0.001 M07 0.056 0.005 0.041 0.002 0.044 0.004 0.0420.004 0.036 0.003 M08 0.006 0.001 0.005 0.000 0.005 0.000 0.006 0.0010.005 0.000 M09 0.019 0.002 0.013 0.001 0.015 0.001 0.014 0.001 0.0120.001 M10 0.054 0.003 0.048 0.001 0.049 0.002 0.047 0.002 0.048 0.003M11 0.105 0.004 0.088 0.003 0.094 0.004 0.090 0.005 0.080 0.004 M120.196 0.012 0.153 0.005 0.166 0.009 0.152 0.010 0.142 0.010 M13 0.0130.001 0.010 0.000 0.011 0.000 0.010 0.001 0.010 0.001 M14 0.046 0.0030.035 0.002 0.040 0.003 0.034 0.002 0.033 0.003 M15 0.073 0.004 0.0590.002 0.061 0.003 0.061 0.003 0.056 0.003 M16 0.250 0.013 0.192 0.0070.214 0.013 0.201 0.012 0.164 0.008 M17 0.408 0.033 0.288 0.015 0.3170.026 0.304 0.026 0.247 0.021 M18 0.024 0.002 0.018 0.001 0.019 0.0010.019 0.002 0.016 0.001 M19 0.103 0.008 0.071 0.004 0.079 0.007 0.0720.006 0.063 0.006 M20 0.077 0.004 0.059 0.002 0.064 0.004 0.060 0.0040.054 0.003 M21 0.265 0.015 0.195 0.008 0.219 0.015 0.205 0.015 0.1650.011 M22 0.933 0.077 0.702 0.036 0.753 0.061 0.738 0.066 0.620 0.057M23 0.018 0.001 0.014 0.001 0.015 0.001 0.014 0.001 0.013 0.001 M240.103 0.009 0.073 0.004 0.079 0.008 0.073 0.007 0.067 0.008 M25 0.2410.013 0.211 0.007 0.218 0.013 0.212 0.014 0.204 0.011 M26 0.069 0.0030.073 0.002 0.076 0.003 0.072 0.004 0.072 0.003

TABLE 25 Ratio and T-test values between females of various levels ofdementia AD, All to CN, Female ADAS 5-19 to CN, Female ADAS 20-39 to CN,Female ADAS 40-70 to CN, Female Metabolite Code Ratio ttest Ratio ttestRatio ttest Ratio ttest M01 0.963 5.2E−01 0.929 2.1E−01 0.951 4.4E−011.004 9.6E−01 M02 0.856 9.0E−02 0.806 4.5E−02 0.912 3.9E−01 0.8331.2E−01 M03 0.772 5.2E−05 0.814 6.0E−03 0.775 8.0E−04 0.737 1.8E−04 M040.963 7.5E−01 0.912 3.8E−01 1.027 8.3E−01 0.932 6.2E−01 M05 0.8931.1E−01 0.925 3.4E−01 0.895 2.1E−01 0.867 8.4E−02 M06 0.843 1.4E−020.910 2.2E−01 0.869 9.8E−02 0.761 5.1E−04 M07 0.732 1.8E−03 0.7985.9E−02 0.753 2.0E−02 0.656 1.1E−03 M08 0.821 5.0E−02 0.849 1.4E−010.889 3.7E−01 0.722 4.7E−03 M09 0.696 2.6E−04 0.777 3.9E−02 0.7022.9E−03 0.624 4.3E−04 M10 0.877 3.1E−02 0.895 8.6E−02 0.860 4.1E−020.883 9.8E−02 M11 0.832 2.5E−03 0.894 7.5E−02 0.850 2.7E−02 0.7621.4E−04 M12 0.778 5.3E−04 0.847 4.2E−02 0.776 4.7E−03 0.726 5.8E−04 M130.800 8.5E−04 0.834 2.5E−02 0.794 8.6E−03 0.780 3.2E−03 M14 0.7724.8E−03 0.869 1.9E−01 0.746 3.0E−03 0.724 5.6E−03 M15 0.811 8.5E−040.835 1.5E−02 0.831 2.3E−02 0.770 4.4E−04 M16 0.765 7.5E−05 0.8534.3E−02 0.801 6.9E−03 0.656 6.4E−08 M17 0.705 3.5E−04 0.776 3.2E−020.745 1.4E−02 0.605 4.3E−05 M18 0.754 3.4E−03 0.804 5.9E−02 0.7926.8E−02 0.673 7.9E−04 M19 0.688 1.8E−04 0.768 2.5E−02 0.699 2.9E−030.612 7.0E−05 M20 0.768 4.9E−04 0.828 3.0E−02 0.782 1.1E−02 0.7034.8E−05 M21 0.737 1.9E−04 0.826 3.7E−02 0.776 1.1E−02 0.624 6.8E−07 M220.752 5.2E−03 0.807 7.0E−02 0.790 6.1E−02 0.665 1.3E−03 M23 0.7642.5E−03 0.809 3.2E−02 0.789 4.1E−02 0.699 7.8E−04 M24 0.708 2.5E−030.768 4.5E−02 0.713 9.3E−03 0.654 4.2E−03 M25 0.876 6.6E−02 0.9072.3E−01 0.880 1.5E−01 0.847 3.5E−02 M26 1.056 3.8E−01 1.092 1.8E−011.043 5.8E−01 1.041 5.5E−01

TABLE 26 Ratio and T-test values between females of various levels ofdementia ADAS 20-39 to 5-19, Female ADAS 40-70 to 5-19, Female ADAS40-70 to 20-39, Female Metabolite Code Ratio ttest Ratio ttest Ratiottest M01 1.023 7.2E−01 1.080 2.8E−01 1.056 4.4E−01 M02 1.132 2.3E−011.034 7.6E−01 0.914 4.1E−01 M03 0.951 5.1E−01 0.906 2.2E−01 0.9525.3E−01 M04 1.126 3.6E−01 1.022 8.8E−01 0.907 5.0E−01 M05 0.967 7.0E−010.936 4.2E−01 0.968 7.1E−01 M06 0.955 5.9E−01 0.837 2.5E−02 0.8761.2E−01 M07 0.944 6.4E−01 0.822 1.2E−01 0.871 2.8E−01 M08 1.046 7.2E−010.850 1.1E−01 0.812 1.1E−01 M09 0.903 3.9E−01 0.802 9.5E−02 0.8893.5E−01 M10 0.961 5.8E−01 0.987 8.6E−01 1.027 7.2E−01 M11 0.951 5.1E−010.853 2.8E−02 0.896 1.6E−01 M12 0.917 3.1E−01 0.858 7.7E−02 0.9354.7E−01 M13 0.953 5.4E−01 0.936 3.6E−01 0.982 8.2E−01 M14 0.858 1.6E−010.833 1.5E−01 0.971 8.0E−01 M15 0.996 9.6E−01 0.922 2.3E−01 0.9263.1E−01 M16 0.940 4.7E−01 0.769 9.7E−04 0.819 1.6E−02 M17 0.960 7.4E−010.779 4.0E−02 0.812 9.8E−02 M18 0.985 9.0E−01 0.837 7.6E−02 0.8501.8E−01 M19 0.911 4.7E−01 0.798 8.5E−02 0.875 3.1E−01 M20 0.945 5.7E−010.849 5.8E−02 0.898 2.7E−01 M21 0.939 5.5E−01 0.755 4.6E−03 0.8044.1E−02 M22 0.979 8.7E−01 0.824 1.2E−01 0.841 1.9E−01 M23 0.975 8.3E−010.864 1.4E−01 0.886 3.0E−01 M24 0.928 6.0E−01 0.852 3.2E−01 0.9185.8E−01 M25 0.971 7.5E−01 0.934 4.0E−01 0.962 6.6E−01 M26 0.955 5.2E−010.954 4.5E−01 0.998 9.8E−01

TABLE 27 Average Serum Ethanolamine Phospholipid Levels in Males ofDifferent Levels of Dementia Severity Cognitive Normal, Male SDAT_all,Male SDAT, ADAS 5-19, Male SDAT, ADAS 20-39, Male SDAT, ADAS 40-70, MaleMetabolite Code Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM M01 0.1160.006 0.119 0.004 0.117 0.006 0.117 0.006 0.117 0.011 M02 0.066 0.0060.056 0.004 0.060 0.009 0.054 0.004 0.046 0.006 M03 0.109 0.007 0.0840.004 0.091 0.006 0.080 0.005 0.072 0.007 M04 0.030 0.002 0.026 0.0020.025 0.004 0.027 0.002 0.022 0.003 M05 0.012 0.001 0.010 0.000 0.0110.001 0.010 0.001 0.010 0.001 M06 0.028 0.002 0.025 0.001 0.024 0.0020.025 0.002 0.024 0.002 M07 0.062 0.005 0.044 0.002 0.047 0.004 0.0440.003 0.036 0.004 M08 0.007 0.001 0.005 0.000 0.006 0.001 0.005 0.0000.004 0.001 M09 0.022 0.002 0.015 0.001 0.016 0.002 0.015 0.001 0.0110.001 M10 0.054 0.002 0.052 0.002 0.053 0.003 0.051 0.002 0.047 0.005M11 0.110 0.005 0.095 0.004 0.095 0.007 0.095 0.006 0.088 0.009 M120.205 0.011 0.162 0.006 0.170 0.011 0.160 0.008 0.141 0.013 M13 0.0130.001 0.011 0.000 0.011 0.001 0.010 0.001 0.010 0.001 M14 0.051 0.0040.036 0.002 0.041 0.004 0.034 0.002 0.028 0.003 M15 0.076 0.004 0.0640.002 0.067 0.005 0.062 0.003 0.060 0.006 M16 0.266 0.015 0.207 0.0090.213 0.015 0.202 0.012 0.191 0.018 M17 0.470 0.038 0.302 0.015 0.3380.031 0.292 0.020 0.242 0.026 M18 0.026 0.003 0.019 0.001 0.020 0.0020.018 0.001 0.016 0.002 M19 0.127 0.013 0.078 0.004 0.088 0.009 0.0760.006 0.056 0.006 M20 0.078 0.005 0.064 0.003 0.069 0.005 0.061 0.0030.059 0.007 M21 0.265 0.017 0.217 0.010 0.226 0.019 0.211 0.014 0.2010.023 M22 1.040 0.087 0.736 0.037 0.789 0.071 0.723 0.052 0.624 0.067M23 0.018 0.001 0.015 0.001 0.016 0.001 0.014 0.001 0.013 0.001 M240.116 0.012 0.079 0.005 0.090 0.010 0.075 0.006 0.061 0.008 M25 0.2400.017 0.218 0.009 0.249 0.018 0.208 0.010 0.183 0.017 M26 0.072 0.0030.070 0.002 0.074 0.004 0.070 0.003 0.065 0.006

TABLE 28 Ratio and T-test Values Between Males of Various Levels ofDementia AD, All to CN, Male ADAS 5-19 to CN, Male ADAS 20-39 to CN,Male ADAS 40-70 to CN, Male Metabolite Code Ratio ttest Ratio ttestRatio ttest Ratio ttest M01 1.028 7.1E−01 1.011 8.9E−01 1.016 8.3E−011.014 8.9E−01 M02 0.845 2.1E−01 0.898 5.5E−01 0.816 7.5E−02 0.6902.4E−02 M03 0.769 1.6E−03 0.836 6.9E−02 0.735 6.4E−04 0.655 1.4E−03 M040.882 3.9E−01 0.836 2.9E−01 0.896 4.1E−01 0.726 2.7E−02 M05 0.8821.3E−01 0.896 2.7E−01 0.865 9.1E−02 0.836 1.5E−01 M06 0.877 1.3E−010.855 9.1E−02 0.876 1.6E−01 0.856 1.4E−01 M07 0.709 3.9E−04 0.7531.8E−02 0.704 1.6E−03 0.589 1.1E−03 M08 0.759 1.9E−02 0.806 1.8E−010.755 2.1E−02 0.629 1.5E−02 M09 0.658 2.7E−04 0.731 2.7E−02 0.6541.4E−03 0.472 2.2E−04 M10 0.960 6.0E−01 0.978 7.9E−01 0.942 4.0E−010.877 1.7E−01 M11 0.866 9.3E−02 0.861 1.1E−01 0.861 8.1E−02 0.7982.6E−02 M12 0.793 1.8E−03 0.831 3.9E−02 0.783 2.2E−03 0.686 8.3E−04 M130.850 2.7E−02 0.890 1.8E−01 0.827 1.3E−02 0.786 1.8E−02 M14 0.7043.7E−04 0.800 6.9E−02 0.666 1.1E−04 0.553 2.3E−04 M15 0.842 2.2E−020.883 1.8E−01 0.810 3.0E−03 0.784 1.9E−02 M16 0.778 1.6E−03 0.8021.7E−02 0.759 1.2E−03 0.718 3.2E−03 M17 0.644 4.5E−06 0.718 7.5E−030.621 1.3E−05 0.515 1.0E−04 M18 0.708 1.6E−03 0.758 6.0E−02 0.6892.0E−03 0.613 7.7E−03 M19 0.611 1.0E−05 0.688 1.1E−02 0.596 6.1E−050.442 2.0E−04 M20 0.826 2.3E−02 0.892 2.7E−01 0.786 6.1E−03 0.7623.9E−02 M21 0.818 2.7E−02 0.852 1.3E−01 0.796 2.0E−02 0.757 2.9E−02 M220.708 4.4E−04 0.758 2.7E−02 0.695 1.3E−03 0.600 1.9E−03 M23 0.8011.1E−02 0.857 1.6E−01 0.776 4.0E−03 0.723 8.6E−03 M24 0.680 1.1E−030.777 9.8E−02 0.647 1.3E−03 0.524 2.8E−03 M25 0.909 2.4E−01 1.0357.4E−01 0.865 8.0E−02 0.760 3.0E−02 M26 0.978 7.4E−01 1.023 7.5E−010.968 6.5E−01 0.907 2.9E−01

TABLE 29 Ratio and T-test Values Between Males of Various Levels ofDementia ADAS 20-39 to 5-19, Male ADAS 40-70 to 5-19, Male ADAS 40-70 to20-39, Male Metabolite Code Ratio ttest Ratio ttest Ratio ttest M011.005 9.4E−01 1.003 9.8E−01 0.998 9.8E−01 M02 0.908 5.3E−01 0.7683.2E−01 0.846 2.8E−01 M03 0.880 1.5E−01 0.784 7.0E−02 0.891 3.4E−01 M041.072 6.8E−01 0.868 5.8E−01 0.810 2.8E−01 M05 0.966 7.0E−01 0.9336.1E−01 0.966 7.7E−01 M06 1.024 8.1E−01 1.001 9.9E−01 0.978 8.6E−01 M070.935 5.5E−01 0.783 1.3E−01 0.837 2.5E−01 M08 0.936 6.3E−01 0.7802.7E−01 0.833 2.4E−01 M09 0.895 4.2E−01 0.646 3.3E−02 0.721 1.0E−01 M100.963 6.3E−01 0.897 3.7E−01 0.932 5.0E−01 M11 1.000 1.0E+00 0.9275.7E−01 0.927 5.4E−01 M12 0.942 4.8E−01 0.826 1.3E−01 0.877 2.4E−01 M130.930 3.8E−01 0.883 3.2E−01 0.950 6.3E−01 M14 0.832 1.1E−01 0.6914.5E−02 0.830 2.0E−01 M15 0.917 3.0E−01 0.888 3.7E−01 0.968 7.4E−01 M160.947 5.4E−01 0.895 3.9E−01 0.946 6.4E−01 M17 0.865 1.9E−01 0.7165.6E−02 0.828 1.9E−01 M18 0.909 4.5E−01 0.809 2.8E−01 0.890 4.4E−01 M190.866 2.5E−01 0.642 3.1E−02 0.741 7.4E−02 M20 0.880 1.6E−01 0.8542.7E−01 0.970 8.0E−01 M21 0.934 5.2E−01 0.889 4.4E−01 0.952 7.2E−01 M220.917 4.5E−01 0.791 1.6E−01 0.863 3.3E−01 M23 0.906 3.3E−01 0.8442.8E−01 0.931 5.6E−01 M24 0.833 1.7E−01 0.674 5.9E−02 0.810 2.3E−01 M250.836 3.7E−02 0.735 2.8E−02 0.879 2.1E−01 M26 0.946 4.3E−01 0.8872.3E−01 0.937 5.2E−01

TABLE 30 Effect of Pathology State on Serum Ethanolamine PhospholipidLevels in Males Post Mortem Ctl, Male Post Mortem SDAT Male SDAT vsControl Metabolite Code Mean SEM Mean SEM Ratio ttest M01 0.127 0.0170.089 0.013 0.702 0.091 M02 0.046 0.006 0.026 0.005 0.568 0.022 M030.059 0.006 0.036 0.006 0.610 0.014 M04 0.017 0.004 0.007 0.002 0.4200.024 M05 0.006 0.001 0.004 0.000 0.479 0.019 M06 0.009 0.001 0.0060.001 0.475 0.005 M07 0.012 0.003 0.009 0.001 0.451 0.033 M08 0.0030.001 0.002 0.000 0.410 0.015 M09 0.006 0.002 0.003 0.001 0.269 0.048M10 0.041 0.005 0.036 0.006 0.608 0.019 M11 0.052 0.006 0.041 0.0060.474 0.001 M12 0.094 0.013 0.084 0.012 0.587 0.024 M13 0.009 0.0010.008 0.001 0.576 0.008 M14 0.025 0.005 0.021 0.007 0.429 0.009 M150.045 0.005 0.029 0.004 0.648 0.026 M16 0.092 0.012 0.053 0.007 0.5700.012 M17 0.097 0.021 0.047 0.007 0.489 0.036 M18 0.010 0.001 0.0050.001 0.521 0.004 M19 0.032 0.006 0.014 0.002 0.452 0.011 M20 0.0310.004 0.017 0.002 0.542 0.006 M21 0.072 0.011 0.033 0.006 0.464 0.006M22 0.217 0.040 0.106 0.017 0.486 0.020 M23 0.009 0.001 0.005 0.0010.565 0.005 M24 0.029 0.004 0.013 0.003 0.448 0.007 M25 0.238 0.0230.180 0.026 0.757 0.114 M26 0.073 0.008 0.050 0.006 0.684 0.034

TABLE 31 Effect of Pathology State on Serum Ethanolamine PhospholipidLevels in Females Post Mortem Ctl, Female Post Mortem SDAT FemaleAutopsy AD vs. Control, Female Metabolite Code Mean SEM Mean SEM Ratiottest M01 0.179 0.050 0.124 0.016 0.697 0.300 M02 0.062 0.022 0.0480.011 0.773 0.557 M03 0.070 0.019 0.043 0.005 0.619 0.178 M04 0.0160.005 0.012 0.004 0.775 0.563 M05 0.007 0.002 0.003 0.001 0.508 0.095M06 0.011 0.003 0.004 0.001 0.531 0.060 M07 0.016 0.003 0.005 0.0010.530 0.037 M08 0.003 0.001 0.001 0.000 0.684 0.258 M09 0.005 0.0010.002 0.000 0.566 0.047 M10 0.053 0.015 0.025 0.004 0.672 0.293 M110.064 0.013 0.025 0.004 0.650 0.132 M12 0.114 0.021 0.055 0.008 0.7420.241 M13 0.010 0.002 0.005 0.001 0.865 0.537 M14 0.023 0.004 0.0110.002 0.904 0.792 M15 0.060 0.017 0.034 0.004 0.577 0.151 M16 0.1070.021 0.064 0.008 0.594 0.061 M17 0.113 0.024 0.067 0.009 0.592 0.079M18 0.014 0.004 0.008 0.001 0.579 0.186 M19 0.035 0.007 0.022 0.0040.635 0.132 M20 0.050 0.014 0.021 0.002 0.410 0.042 M21 0.100 0.0200.041 0.007 0.414 0.010 M22 0.283 0.058 0.168 0.024 0.593 0.071 M230.012 0.003 0.007 0.001 0.560 0.112 M24 0.035 0.007 0.018 0.002 0.4960.025 M25 0.189 0.015 0.198 0.022 1.051 0.727 M26 0.054 0.008 0.0610.006 1.123 0.496

TABLE 32 Effect of Age on Ethanolamine Phospholipid Ratios to M01 inMales Age Ctl, 30-39, Male Age Ctl, 40-49, Male Age Ctl, 50-59, Male AgeCtl, 60-69, Male Age Ctl, 70+_Male Metabolite Code Mean SEM Mean SEMMean SEM Mean SEM Mean SEM M01 1.000 0.000 1.000 0.000 1.000 0.000 1.0000.000 1.000 0.000 M02 0.476 0.042 0.477 0.034 0.517 0.027 0.431 0.0310.420 0.028 M03 0.863 0.063 0.751 0.049 0.864 0.030 0.792 0.051 0.8480.071 M04 0.224 0.025 0.229 0.026 0.235 0.013 0.203 0.019 0.204 0.017M05 0.113 0.008 0.102 0.007 0.101 0.004 0.091 0.005 0.088 0.005 M060.263 0.029 0.234 0.021 0.243 0.012 0.199 0.016 0.210 0.012 M07 0.6830.136 0.493 0.070 0.581 0.041 0.479 0.052 0.457 0.039 M08 0.079 0.0100.064 0.007 0.067 0.004 0.054 0.005 0.049 0.004 M09 0.161 0.025 0.1240.017 0.173 0.013 0.154 0.018 0.172 0.024 M10 0.464 0.025 0.454 0.0220.445 0.012 0.419 0.018 0.407 0.016 M11 1.063 0.111 0.899 0.069 0.9080.037 0.789 0.052 0.811 0.038 M12 2.068 0.251 1.728 0.168 1.942 0.0991.664 0.111 1.552 0.087 M13 0.126 0.009 0.117 0.009 0.121 0.005 0.1020.006 0.092 0.005 M14 0.294 0.033 0.242 0.029 0.332 0.020 0.316 0.0290.363 0.041 M15 0.699 0.034 0.658 0.041 0.675 0.023 0.567 0.031 0.5900.029 M16 2.429 0.285 2.020 0.175 2.284 0.118 1.778 0.145 1.922 0.112M17 4.127 0.757 3.164 0.418 3.980 0.257 3.167 0.320 3.178 0.294 M180.258 0.025 0.229 0.025 0.244 0.014 0.195 0.021 0.174 0.014 M19 0.7250.101 0.607 0.094 0.865 0.063 0.769 0.085 0.881 0.131 M20 0.691 0.0490.616 0.043 0.619 0.030 0.529 0.034 0.553 0.034 M21 2.395 0.215 2.1630.189 2.287 0.122 1.773 0.148 1.840 0.108 M22 8.811 1.529 7.346 0.8838.537 0.549 7.124 0.644 6.920 0.635 M23 0.189 0.019 0.175 0.016 0.1780.009 0.150 0.016 0.133 0.008 M24 0.649 0.086 0.606 0.098 0.787 0.0570.700 0.072 0.799 0.122 M25 2.182 0.444 1.975 0.265 2.747 0.211 2.6080.251 3.066 0.284

TABLE 33 Ratio and T-test values of Ethanolamine Phospholipid Ratios toM01 between males of different ages 50-59 vs. 40-49, Male 60-69 vs.40-49, Male 70+ vs. 40-49, Male Metabolite Code Ratio ttest Ratio ttestRatio ttest M01 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 1.0823.9E−01 0.903 3.2E−01 0.879 1.9E−01 M03 1.149 4.5E−02 1.054 5.8E−011.128 3.0E−01 M04 1.023 8.4E−01 0.883 4.0E−01 0.889 3.9E−01 M05 0.9848.3E−01 0.884 1.6E−01 0.863 8.5E−02 M06 1.041 6.8E−01 0.851 1.9E−010.898 3.1E−01 M07 1.177 2.6E−01 0.970 8.6E−01 0.927 6.4E−01 M08 1.0536.7E−01 0.847 2.6E−01 0.765 5.4E−02 M09 1.404 2.5E−02 1.243 2.4E−011.391 1.3E−01 M10 0.981 7.1E−01 0.924 2.3E−01 0.896 7.6E−02 M11 1.0109.0E−01 0.878 2.0E−01 0.903 2.5E−01 M12 1.124 2.5E−01 0.963 7.4E−010.898 3.3E−01 M13 1.035 6.7E−01 0.873 1.7E−01 0.787 1.3E−02 M14 1.3721.2E−02 1.306 7.7E−02 1.500 2.7E−02 M15 1.026 7.0E−01 0.862 7.8E−020.896 1.7E−01 M16 1.131 2.1E−01 0.880 2.9E−01 0.951 6.2E−01 M17 1.2588.8E−02 1.001 1.0E+00 1.004 9.8E−01 M18 1.067 5.6E−01 0.851 3.0E−010.761 4.9E−02 M19 1.426 2.4E−02 1.266 2.1E−01 1.451 1.1E−01 M20 1.0049.6E−01 0.858 1.1E−01 0.898 2.5E−01 M21 1.057 5.8E−01 0.820 1.0E−010.851 1.2E−01 M22 1.162 2.4E−01 0.970 8.4E−01 0.942 6.9E−01 M23 1.0168.7E−01 0.858 2.8E−01 0.759 1.5E−02 M24 1.300 9.4E−02 1.156 4.3E−011.319 2.4E−01 M25 1.391 3.5E−02 1.321 9.0E−02 1.553 8.2E−03

TABLE 34 Effect of Age on Ethanolamine Phospholipid Ratios to M01 inFemales Age Ctl, 30-39, Female Age Ctl, 40-49, Female Age Ctl, 50-59,Female Age Ctl, 60-69, Female Age Ctl, 70+_Female Metabolite Code MeanSEM Mean SEM Mean SEM Mean SEM Mean SEM M01 1.000 0.000 1.000 0.0001.000 0.000 1.000 0.000 1.000 0.000 M02 0.474 0.037 0.597 0.028 0.5510.020 0.460 0.018 0.455 0.023 M03 0.931 0.089 0.911 0.043 0.926 0.0300.750 0.034 0.838 0.048 M04 0.279 0.036 0.283 0.017 0.241 0.011 0.1840.010 0.221 0.020 M05 0.112 0.009 0.118 0.006 0.105 0.003 0.085 0.0050.089 0.006 M06 0.311 0.044 0.291 0.019 0.261 0.010 0.193 0.012 0.2130.017 M07 0.618 0.090 0.667 0.056 0.582 0.030 0.435 0.039 0.430 0.041M08 0.079 0.008 0.089 0.007 0.070 0.003 0.047 0.004 0.052 0.007 M090.204 0.036 0.194 0.017 0.197 0.011 0.139 0.012 0.163 0.015 M10 0.4910.027 0.535 0.024 0.490 0.010 0.424 0.015 0.422 0.022 M11 1.220 0.1591.092 0.060 1.021 0.031 0.823 0.037 0.840 0.058 M12 1.979 0.150 2.0440.117 1.952 0.063 1.534 0.086 1.564 0.100 M13 0.129 0.008 0.136 0.0070.122 0.004 0.092 0.005 0.104 0.010 M14 0.379 0.047 0.381 0.024 0.4140.018 0.333 0.021 0.412 0.033 M15 0.706 0.044 0.720 0.034 0.699 0.0190.560 0.024 0.578 0.033 M16 2.739 0.333 2.483 0.161 2.345 0.091 1.7830.096 1.964 0.136 M17 4.055 0.499 4.149 0.401 3.987 0.209 2.963 0.2753.010 0.268 M18 0.282 0.022 0.289 0.022 0.245 0.010 0.160 0.012 0.1910.024 M19 1.009 0.175 0.902 0.085 0.974 0.056 0.716 0.061 0.877 0.077M20 0.708 0.059 0.682 0.045 0.637 0.021 0.524 0.029 0.565 0.039 M212.885 0.411 2.537 0.187 2.331 0.092 1.757 0.105 1.935 0.158 M22 8.9710.932 9.244 0.790 8.625 0.450 6.263 0.484 6.923 0.683 M23 0.198 0.0110.202 0.014 0.177 0.006 0.124 0.008 0.140 0.014 M24 0.955 0.174 0.8340.078 0.882 0.049 0.661 0.054 0.814 0.071 M25 2.505 0.217 2.286 0.1643.219 0.181 2.995 0.239 3.280 0.248

TABLE 35 Ratio and T-test values of Ethanolamine Phospholipid Ratios toM01 between females of different ages 50-59 vs. 40-49, Female 60-69 vs.40-49, Female 70+ vs. 40-49, Female Metabolite Code Ratio ttest Ratiottest Ratio ttest M01 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M020.923 2.1E−01 0.771 5.7E−05 0.763 8.4E−04 M03 1.017 7.8E−01 0.8233.7E−03 0.920 2.8E−01 M04 0.850 3.5E−02 0.652 1.1E−06 0.782 2.3E−02 M050.892 4.7E−02 0.721 5.0E−05 0.758 3.7E−03 M06 0.896 1.3E−01 0.6621.3E−05 0.733 6.5E−03 M07 0.873 1.5E−01 0.652 7.2E−04 0.644 3.5E−03 M080.783 4.7E−03 0.533 6.9E−07 0.583 9.9E−04 M09 1.016 8.8E−01 0.7177.8E−03 0.841 2.2E−01 M10 0.917 4.8E−02 0.792 7.9E−05 0.789 2.0E−03 M110.935 2.5E−01 0.754 1.3E−04 0.770 5.9E−03 M12 0.955 4.6E−01 0.7515.1E−04 0.765 5.8E−03 M13 0.900 7.0E−02 0.680 4.8E−06 0.764 9.1E−03 M141.088 3.1E−01 0.876 1.4E−01 1.083 4.3E−01 M15 0.970 5.6E−01 0.7781.8E−04 0.802 6.7E−03 M16 0.944 4.3E−01 0.718 1.8E−04 0.791 2.8E−02 M170.961 7.0E−01 0.714 1.4E−02 0.726 4.4E−02 M18 0.847 4.1E−02 0.5526.0E−07 0.660 5.6E−03 M19 1.079 4.9E−01 0.794 7.2E−02 0.972 8.4E−01 M200.935 3.1E−01 0.769 2.9E−03 0.828 7.5E−02 M21 0.919 2.7E−01 0.6932.4E−04 0.763 2.9E−02 M22 0.933 4.8E−01 0.678 1.2E−03 0.749 4.6E−02 M230.876 5.5E−02 0.614 1.3E−06 0.696 3.9E−03 M24 1.058 6.0E−01 0.7936.5E−02 0.976 8.6E−01 M25 1.408 2.4E−03 1.310 2.2E−02 1.435 8.6E−04

TABLE 36 Average Serum Ethanolamine Phospholipid Ratios to M01 in Malesof Different Levels of Dementia Severity Cognitive Normal, MaleSDAT_all, Male SDAT, ADAS 5-19, Male SDAT, ADAS 20-39, Male SDAT, ADAS40-70, Male Metabolite Code Mean SEM Mean SEM Mean SEM Mean SEM Mean SEMM01 1.000 0.000 1.000 0.000 1.000 0.000 1.000 0.000 1.000 0.000 M020.564 0.031 0.453 0.015 0.475 0.032 0.455 0.017 0.383 0.022 M03 0.9600.050 0.712 0.018 0.779 0.037 0.694 0.024 0.617 0.027 M04 0.262 0.0150.210 0.009 0.203 0.017 0.219 0.013 0.183 0.014 M05 0.106 0.006 0.0890.002 0.091 0.004 0.089 0.003 0.085 0.004 M06 0.255 0.014 0.213 0.0070.211 0.012 0.214 0.010 0.211 0.012 M07 0.554 0.041 0.373 0.014 0.3960.025 0.375 0.020 0.321 0.028 M08 0.062 0.005 0.044 0.002 0.047 0.0040.045 0.003 0.038 0.003 M09 0.195 0.015 0.126 0.006 0.139 0.012 0.1270.009 0.093 0.008 M10 0.483 0.021 0.440 0.009 0.450 0.016 0.441 0.0130.408 0.016 M11 0.985 0.043 0.817 0.025 0.828 0.051 0.824 0.035 0.7610.036 M12 1.843 0.088 1.399 0.036 1.460 0.059 1.406 0.053 1.252 0.090M13 0.114 0.006 0.092 0.002 0.096 0.004 0.091 0.003 0.088 0.006 M140.442 0.023 0.310 0.013 0.348 0.028 0.301 0.015 0.255 0.025 M15 0.6820.031 0.548 0.012 0.571 0.022 0.541 0.016 0.520 0.024 M16 2.398 0.1281.790 0.057 1.856 0.105 1.777 0.082 1.687 0.115 M17 4.203 0.304 2.5690.105 2.853 0.205 2.501 0.135 2.187 0.243 M18 0.232 0.017 0.156 0.0060.166 0.012 0.155 0.009 0.140 0.010 M19 1.103 0.092 0.663 0.032 0.7400.066 0.660 0.041 0.503 0.045 M20 0.692 0.037 0.548 0.016 0.593 0.0300.532 0.021 0.509 0.034 M21 2.377 0.126 1.857 0.066 1.951 0.128 1.8290.090 1.754 0.143 M22 9.309 0.674 6.230 0.231 6.651 0.434 6.157 0.3055.616 0.592 M23 0.164 0.009 0.125 0.004 0.133 0.008 0.124 0.006 0.1140.008 M24 1.010 0.088 0.672 0.034 0.770 0.076 0.647 0.039 0.539 0.053M25 2.160 0.133 2.085 0.115 2.215 0.145 2.102 0.197 1.811 0.222

TABLE 37 Ratio and T-test Values of Ethanolamine Phospholipid Ratios toM01 between Males of Various Levels of Dementia AD, All to CN, Male ADAS5-19 to CN, Male ADAS 20-39 to CN, Male ADAS 40-70 to CN, MaleMetabolite Code Ratio ttest Ratio ttest Ratio ttest Ratio ttest M011.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 0.8037.2E−04 0.842 5.2E−02 0.807 1.2E−03 0.680 1.8E−04 M03 0.742 6.4E−080.812 3.8E−03 0.724 4.5E−07 0.643 9.8E−06 M04 0.802 8.4E−03 0.7741.6E−02 0.836 4.1E−02 0.700 1.3E−03 M05 0.840 9.2E−04 0.858 2.9E−020.841 5.5E−03 0.798 1.2E−02 M06 0.833 5.0E−03 0.827 2.0E−02 0.8402.2E−02 0.828 4.7E−02 M07 0.673 4.7E−07 0.715 1.1E−03 0.676 3.1E−050.580 2.4E−04 M08 0.713 1.1E−04 0.747 1.3E−02 0.727 1.5E−03 0.6028.4E−04 M09 0.645 2.8E−06 0.713 4.4E−03 0.654 7.5E−05 0.476 1.6E−05 M100.910 3.4E−02 0.932 2.1E−01 0.913 8.0E−02 0.843 1.8E−02 M11 0.8301.9E−03 0.841 2.5E−02 0.837 5.8E−03 0.773 9.3E−04 M12 0.759 3.4E−070.792 3.9E−04 0.763 1.9E−05 0.679 7.0E−05 M13 0.811 1.0E−04 0.8461.2E−02 0.802 4.0E−04 0.772 4.7E−03 M14 0.701 4.1E−06 0.787 1.4E−020.680 1.1E−06 0.576 4.9E−06 M15 0.803 2.7E−06 0.837 3.8E−03 0.7922.4E−05 0.762 8.6E−04 M16 0.747 4.3E−06 0.774 1.5E−03 0.741 4.8E−050.704 5.4E−04 M17 0.611 2.1E−09 0.679 3.1E−04 0.595 7.2E−08 0.5204.0E−05 M18 0.672 9.8E−07 0.714 1.7E−03 0.666 2.3E−05 0.603 3.3E−04 M190.601 6.5E−08 0.672 1.7E−03 0.599 2.1E−06 0.456 2.2E−05 M20 0.7939.1E−05 0.858 3.8E−02 0.769 1.0E−04 0.736 1.8E−03 M21 0.782 3.7E−040.821 2.2E−02 0.770 5.6E−04 0.738 3.0E−03 M22 0.669 2.0E−07 0.7149.9E−04 0.661 4.8E−06 0.603 6.0E−04 M23 0.763 1.2E−04 0.810 1.5E−020.754 4.8E−04 0.696 6.5E−04 M24 0.665 3.5E−05 0.763 4.2E−02 0.6413.5E−05 0.533 4.0E−04 M25 0.965 7.5E−01 1.025 7.9E−01 0.973 8.4E−010.838 1.6E−01

TABLE 38 Ratio and T-test Values of Ethanolamine Phospholipid Ratios toM01 Between Males of Various Levels of Dementia ADAS 20-39 to 5-19, MaleADAS 40-70 to 5-19, Male ADAS 40-70 to 20-39, Male Metabolite Code Ratiottest Ratio ttest Ratio ttest M01 1.000 #DIV/0! 1.000 #DIV/0! 1.000#DIV/0! M02 0.958 5.5E−01 0.807 7.1E−02 0.843 3.5E−02 M03 0.891 4.6E−020.792 6.9E−03 0.888 9.3E−02 M04 1.080 4.5E−01 0.903 4.8E−01 0.8371.4E−01 M05 0.980 7.2E−01 0.930 3.2E−01 0.948 4.4E−01 M06 1.016 8.4E−011.002 9.8E−01 0.987 8.9E−01 M07 0.946 5.1E−01 0.811 8.4E−02 0.8571.8E−01 M08 0.973 7.8E−01 0.806 1.3E−01 0.828 1.3E−01 M09 0.917 4.2E−010.668 1.5E−02 0.728 3.5E−02 M10 0.980 6.6E−01 0.905 1.1E−01 0.9231.9E−01 M11 0.995 9.4E−01 0.919 4.0E−01 0.924 3.5E−01 M12 0.964 5.1E−010.858 5.8E−02 0.890 1.6E−01 M13 0.948 3.5E−01 0.913 2.6E−01 0.9636.3E−01 M14 0.864 1.1E−01 0.732 4.1E−02 0.847 1.4E−01 M15 0.947 2.6E−010.911 1.7E−01 0.962 5.2E−01 M16 0.957 5.5E−01 0.909 3.4E−01 0.9505.8E−01 M17 0.877 1.4E−01 0.767 6.0E−02 0.875 2.6E−01 M18 0.932 4.5E−010.844 1.9E−01 0.905 3.9E−01 M19 0.891 2.8E−01 0.679 2.6E−02 0.7624.5E−02 M20 0.896 8.6E−02 0.858 9.6E−02 0.957 5.9E−01 M21 0.937 4.2E−010.899 3.6E−01 0.959 6.8E−01 M22 0.926 3.4E−01 0.844 1.8E−01 0.9124.0E−01 M23 0.931 3.9E−01 0.860 1.8E−01 0.923 4.5E−01 M24 0.840 1.2E−010.699 5.7E−02 0.832 1.5E−01 M25 0.949 6.7E−01 0.818 1.3E−01 0.8624.4E−01

TABLE 39 Effect of Dementia State on Ethanolamine Phospholipid Ratios toM01 in Females Cognitive Normal, SDAT_all, SDAT, ADAS 5-19, SDAT, ADAS20-39, SDAT, ADAS 40-70, Female Female Female Female Female MetaboliteCode Mean SEM Mean SEM Mean SEM Mean SEM Mean SEM M01 1.000 0.000 1.0000.000 1.000 0.000 1.000 0.000 1.000 0.000 M02 0.520 0.029 0.463 0.0120.454 0.021 0.499 0.023 0.431 0.018 M03 0.852 0.034 0.687 0.017 0.7420.030 0.704 0.031 0.624 0.022 M04 0.222 0.013 0.219 0.010 0.215 0.0130.241 0.021 0.197 0.015 M05 0.092 0.004 0.086 0.003 0.091 0.004 0.0880.005 0.081 0.004 M06 0.234 0.012 0.207 0.007 0.226 0.011 0.217 0.0130.180 0.009 M07 0.474 0.034 0.368 0.018 0.422 0.039 0.377 0.028 0.3140.025 M08 0.054 0.004 0.047 0.002 0.050 0.003 0.051 0.004 0.039 0.002M09 0.167 0.013 0.121 0.005 0.140 0.010 0.123 0.009 0.103 0.008 M100.469 0.016 0.431 0.010 0.451 0.016 0.428 0.018 0.418 0.016 M11 0.9290.039 0.807 0.023 0.886 0.041 0.839 0.044 0.708 0.030 M12 1.682 0.0781.384 0.042 1.568 0.090 1.384 0.067 1.239 0.062 M13 0.111 0.005 0.0950.003 0.101 0.005 0.095 0.005 0.089 0.004 M14 0.392 0.023 0.315 0.0130.367 0.029 0.309 0.019 0.279 0.019 M15 0.627 0.024 0.539 0.014 0.5640.021 0.563 0.030 0.493 0.016 M16 2.214 0.114 1.780 0.064 1.994 0.1071.906 0.127 1.470 0.069 M17 3.497 0.247 2.593 0.127 2.976 0.266 2.7500.226 2.115 0.146 M18 0.202 0.014 0.161 0.007 0.178 0.011 0.170 0.0140.138 0.008 M19 0.895 0.061 0.634 0.030 0.728 0.057 0.656 0.055 0.5370.041 M20 0.669 0.035 0.535 0.018 0.587 0.032 0.553 0.036 0.474 0.022M21 2.318 0.118 1.798 0.076 2.039 0.132 1.922 0.147 1.467 0.086 M228.068 0.596 6.310 0.304 7.041 0.600 6.681 0.568 5.313 0.360 M23 0.1580.009 0.127 0.005 0.139 0.009 0.131 0.010 0.112 0.006 M24 0.893 0.0680.651 0.034 0.724 0.063 0.669 0.059 0.573 0.053 M25 2.145 0.127 2.0310.086 2.072 0.129 2.109 0.162 1.910 0.139

TABLE 40 Ratio and T-test values of Ethanolamine Phospholipid Ratios toM01 between females of various levels of dementia AD, All to CN, FemaleADAS 5-19 to CN, Female ADAS 20-39 to CN, Female ADAS 40-70 to CN,Female Metabolite Code Ratio ttest Ratio ttest Ratio ttest Ratio ttestM01 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! 1.000 #DIV/0! M02 0.8914.7E−02 0.872 6.4E−02 0.960 5.7E−01 0.828 7.0E−03 M03 0.806 1.3E−050.871 1.6E−02 0.826 2.1E−03 0.732 6.8E−08 M04 0.986 8.8E−01 0.9687.0E−01 1.086 4.9E−01 0.887 2.3E−01 M05 0.941 3.5E−01 0.989 8.7E−010.962 6.4E−01 0.879 5.5E−02 M06 0.885 6.6E−02 0.966 6.3E−01 0.9283.7E−01 0.771 2.8E−04 M07 0.776 6.7E−03 0.890 3.2E−01 0.796 3.2E−020.663 2.3E−04 M08 0.869 1.3E−01 0.934 4.9E−01 0.943 6.2E−01 0.7351.5E−03 M09 0.724 3.1E−04 0.837 1.0E−01 0.740 5.4E−03 0.618 4.2E−05 M100.919 7.3E−02 0.963 4.4E−01 0.913 1.1E−01 0.892 3.1E−02 M11 0.8681.6E−02 0.953 4.4E−01 0.902 1.5E−01 0.762 2.0E−05 M12 0.823 1.5E−030.932 3.4E−01 0.823 5.0E−03 0.736 2.0E−05 M13 0.854 6.6E−03 0.9141.8E−01 0.854 3.1E−02 0.806 9.4E−04 M14 0.802 5.5E−03 0.935 4.9E−010.789 6.6E−03 0.711 2.4E−04 M15 0.860 4.8E−03 0.900 5.1E−02 0.8971.3E−01 0.786 9.1E−06 M16 0.804 2.1E−03 0.901 1.6E−01 0.861 9.3E−020.664 8.8E−08 M17 0.742 1.5E−03 0.851 1.6E−01 0.786 3.2E−02 0.6052.3E−06 M18 0.798 9.5E−03 0.879 1.8E−01 0.844 1.4E−01 0.683 6.3E−05 M190.709 1.6E−04 0.813 5.1E−02 0.733 5.3E−03 0.600 3.2E−06 M20 0.8001.0E−03 0.878 8.7E−02 0.826 2.8E−02 0.709 3.4E−06 M21 0.776 1.4E−030.879 1.2E−01 0.829 5.6E−02 0.633 5.4E−08 M22 0.782 9.6E−03 0.8732.3E−01 0.828 1.1E−01 0.659 7.8E−05 M23 0.801 5.3E−03 0.877 1.3E−010.831 6.8E−02 0.707 4.5E−05 M24 0.729 1.6E−03 0.811 7.2E−02 0.7501.7E−02 0.642 3.3E−04 M25 0.947 5.3E−01 0.966 6.9E−01 0.983 8.7E−010.890 2.3E−01

TABLE 41 Ratio and T-test values of Ethanolamine Phospholipid Ratios toM01 between females of various levels of dementia ADAS 20-39 to 5-19,Female ADAS 40-70 to 5-19, Female ADAS 40-70 to 20-39, Female MetaboliteCode Ratio ttest Ratio ttest Ratio ttest M01 1.000 #DIV/0! 1.000 #DIV/0!1.000 #DIV/0! M02 1.101 1.6E−01 0.949 4.1E−01 0.862 2.2E−02 M03 0.9494.0E−01 0.841 1.5E−03 0.887 4.2E−02 M04 1.122 3.3E−01 0.917 3.8E−010.817 9.2E−02 M05 0.972 7.3E−01 0.889 7.6E−02 0.914 2.6E−01 M06 0.9616.3E−01 0.797 1.3E−03 0.830 2.4E−02 M07 0.895 3.4E−01 0.745 1.7E−020.833 9.8E−02 M08 1.010 9.4E−01 0.787 1.1E−02 0.780 3.5E−02 M09 0.8842.4E−01 0.738 6.7E−03 0.835 1.0E−01 M10 0.948 3.5E−01 0.926 1.5E−010.977 6.8E−01 M11 0.947 4.6E−01 0.799 6.4E−04 0.844 1.9E−02 M12 0.8839.7E−02 0.790 2.5E−03 0.895 1.2E−01 M13 0.934 3.6E−01 0.882 5.3E−020.944 4.1E−01 M14 0.844 8.6E−02 0.761 9.9E−03 0.902 2.6E−01 M15 0.9989.7E−01 0.874 7.7E−03 0.876 5.3E−02 M16 0.956 6.2E−01 0.737 5.0E−050.771 4.2E−03 M17 0.924 5.2E−01 0.711 3.6E−03 0.769 2.4E−02 M18 0.9607.2E−01 0.777 4.1E−03 0.809 5.7E−02 M19 0.901 3.8E−01 0.738 7.1E−030.819 9.2E−02 M20 0.941 4.9E−01 0.808 3.7E−03 0.858 7.3E−02 M21 0.9435.8E−01 0.720 3.1E−04 0.763 1.1E−02 M22 0.949 6.7E−01 0.755 1.2E−020.795 5.1E−02 M23 0.948 6.1E−01 0.806 1.4E−02 0.851 1.1E−01 M24 0.9255.4E−01 0.791 6.9E−02 0.856 2.3E−01 M25 1.018 8.7E−01 0.922 4.0E−010.906 3.6E−01

TABLE 42 Effect of Pathology State on Ethanolamine Phospholipid Ratiosto M01 in Males Post Mortem Ctl, Male Post Mortem SDAT Male SDAT vsControl Metabolite Code Mean SEM Mean SEM Ratio ttest M01 1.000 0.0001.000 0.000 1.000 #DIV/0! M02 0.367 0.029 0.290 0.029 0.791 0.076 M030.482 0.042 0.391 0.013 0.811 0.054 M04 0.143 0.031 0.076 0.009 0.5290.052 M05 0.048 0.009 0.029 0.002 0.607 0.052 M06 0.080 0.014 0.0460.003 0.581 0.033 M07 0.107 0.025 0.059 0.005 0.549 0.074 M08 0.0240.005 0.012 0.001 0.511 0.037 M09 0.052 0.018 0.018 0.002 0.341 0.074M10 0.337 0.038 0.269 0.013 0.798 0.107 M11 0.452 0.066 0.272 0.0170.602 0.016 M12 0.819 0.130 0.616 0.024 0.753 0.143 M13 0.079 0.0090.060 0.005 0.759 0.085 M14 0.212 0.039 0.115 0.010 0.542 0.026 M150.375 0.035 0.344 0.025 0.918 0.483 M16 0.792 0.128 0.627 0.047 0.7910.240 M17 0.849 0.191 0.561 0.040 0.660 0.156 M18 0.085 0.011 0.0560.004 0.656 0.018 M19 0.273 0.051 0.164 0.015 0.601 0.056 M20 0.2540.027 0.187 0.009 0.737 0.028 M21 0.613 0.109 0.371 0.021 0.605 0.042M22 1.890 0.379 1.199 0.083 0.634 0.092 M23 0.074 0.008 0.055 0.0040.750 0.061 M24 0.241 0.039 0.142 0.011 0.588 0.025 M25 2.066 0.2252.384 0.388 1.154 0.487

TABLE 43 Effect of Pathology State on Ethanolamine Phospholipid Ratiosto M01 in Females Post Mortem Ctl, Female Post Mortem SDAT FemaleAutopsy AD vs. Control, Female Metabolite Code Mean SEM Mean SEM Ratiottest M01 1.000 0.000 1.000 0.000 1.000 #DIV/0! M02 0.322 0.028 0.3670.047 1.140 0.440 M03 0.400 0.022 0.361 0.023 0.902 0.237 M04 0.0860.012 0.092 0.019 1.069 0.798 M05 0.040 0.003 0.032 0.005 0.809 0.225M06 0.069 0.008 0.059 0.017 0.855 0.605 M07 0.102 0.016 0.077 0.0130.752 0.242 M08 0.017 0.002 0.017 0.002 0.981 0.922 M09 0.033 0.0050.024 0.003 0.725 0.133 M10 0.290 0.025 0.291 0.027 1.003 0.981 M110.384 0.044 0.364 0.057 0.950 0.797 M12 0.731 0.096 0.699 0.059 0.9570.777 M13 0.061 0.008 0.069 0.007 1.122 0.489 M14 0.144 0.015 0.1550.033 1.071 0.791 M15 0.339 0.021 0.301 0.027 0.889 0.297 M16 0.6700.066 0.599 0.112 0.895 0.605 M17 0.697 0.063 0.591 0.082 0.848 0.329M18 0.081 0.014 0.069 0.008 0.854 0.462 M19 0.211 0.017 0.180 0.0180.853 0.230 M20 0.283 0.025 0.193 0.032 0.680 0.042 M21 0.624 0.0790.421 0.127 0.674 0.203 M22 1.782 0.251 1.480 0.225 0.831 0.382 M230.073 0.011 0.060 0.006 0.816 0.275 M24 0.214 0.018 0.149 0.016 0.6970.016 M25 1.382 0.214 1.767 0.221 1.279 0.229

TABLE 44 Effect of Dementia State on White and Gray Matter Scores inMales White Matter Gray Matter Score Score Mean SEM Mean SEM CohortCognitive Normal, Male −0.25 0.08 −0.44 0.11 SDAT_all, Male −0.63 0.06−1.11 0.08 SDAT, ADAS 5-19, Male −0.56 0.09 −1.00 0.13 SDAT, ADAS 20-39,Male −0.67 0.08 −1.11 0.10 SDAT, ADAS 40-70, Male −0.71 0.14 −1.45 0.16Post Mortem Ctl, Male −0.44 0.13 −0.41 0.21 Post Mortem SDAT Male −1.620.19 −1.28 0.23 Comparison AD, All to CN, Male Delta −0.37 Delta −0.67ttest 1.9E−03 ttest 2.4E−05 ADAS 5-19 to CN, Male Delta −0.31 Delta−0.56 ttest 1.8E−02 ttest 2.5E−03 ADAS 20-39 to CN, Male Delta −0.41Delta −0.67 ttest 1.5E−03 ttest 7.8E−05 ADAS 40-70 to CN, Male Delta−0.46 Delta −1.01 ttest 3.7E−03 ttest 1.8E−06 ADAS 20-39 to 5-19, MaleDelta −0.10 Delta −0.11 ttest 4.2E−01 ttest 5.0E−01 ADAS 40-70 to 5-19,Male Delta −0.15 Delta −0.45 ttest 3.9E−01 ttest 5.2E−02 ADAS 40-70 to20-39, Male Delta −0.04 Delta −0.34 ttest 8.0E−01 ttest 1.1E−01 AutopsyAD vs. Control, Male Delta −0.89 Delta −1.13 ttest 8.9E−03 ttest 2.8E−03

TABLE 45 Effect of Dementia State on White and Gray Matter Scores inFemales White Matter Gray Matter Score Score Mean SEM Mean SEM CohortCognitive Normal, Female −0.27 0.09 −0.42 0.13 SDAT_all, Female −0.730.05 −1.01 0.07 SDAT, ADAS 5-19, Female −0.55 0.08 −0.85 0.11 SDAT, ADAS20-39, Female −0.69 0.09 −0.94 0.11 SDAT, ADAS 40-70, Female −0.91 0.08−1.21 0.12 Post Mortem Ctl, Female −0.50 0.29 −0.53 0.26 Post MortemSDAT Female −1.34 0.27 −1.54 0.25 Comparison AD, All to CN, Female Delta−0.46 Delta −0.59 ttest 4.6E−05 ttest 1.1E−04 ADAS 5-19 to CN, FemaleDelta −0.29 Delta −0.43 ttest 1.7E−02 ttest 1.3E−02 ADAS 20-39 to CN,Female Delta −0.43 Delta −0.52 ttest 1.9E−03 ttest 3.6E−03 ADAS 40-70 toCN, Female Delta −0.64 Delta −0.79 ttest 9.5E−07 ttest 2.2E−05 ADAS20-39 to 5-19, Female Delta −0.14 Delta −0.09 ttest 2.7E−01 ttest5.8E−01 ADAS 40-70 to 5-19, Female Delta −0.36 Delta −0.37 ttest 2.9E−03ttest 3.1E−02 ADAS 40-70 to 20-39, Female Delta −0.21 Delta −0.27 ttest9.4E−02 ttest 1.0E−01 Autopsy AD vs. Control, Female Delta −1.17 Delta−0.81 ttest 2.0E−03 ttest 2.5E−02

TABLE 46 Distribution of White and Gray Matter Scores in Males (MeanNormalized to CN Male) ADAS-cog ADAS-cog ADAS-cog Autopsy Autopsy MMSE ≧28 8-19 20-39 40-70 Control AD Bin Frequency Frequency FrequencyFrequency Frequency Frequency White Matter Distribution −2 0 1 1 0 0 2−1.75 0 0 0 0 0 0 −1.5 0 0 3 1 0 0 −1.25 0 2 4 0 0 1 −1 1 2 3 1 0 2−0.75 1 3 3 4 0 2 −0.5 3 2 6 4 2 0 −0.25 3 8 13 4 3 1 0 7 13 12 0 2 10.25 9 6 5 1 2 1 0.5 5 1 5 2 0 0 0.75 1 0 1 0 1 0 1 2 0 2 1 0 0 More 0 20 0 0 0 Gray Matter distribution −2 0 1 4 2 0 2 −1.75 0 2 2 1 0 0 −1.5 01 3 0 0 0 −1.25 1 4 2 2 0 1 −1 0 6 7 4 0 2 −0.75 1 4 9 2 1 3 −0.5 7 3 54 1 1 −0.25 3 5 9 1 2 0 0 3 2 7 1 1 0 0.25 5 4 2 1 3 0 0.5 7 3 4 0 1 10.75 3 1 2 0 0 0 1 0 3 2 0 0 0 More 2 1 0 0 1 0

TABLE 47 Distribution of White and Gray Matter Scores in Females (MeanNormalized to CN Female) ADAS-cog ADAS-cog ADAS-cog Autopsy Autopsy MMSE≧ 28 8-19 20-39 40-70 Control AD Bin Frequency Frequency FrequencyFrequency Frequency Frequency White Matter Distribution −2 0 0 0 1 0 1−1.75 0 0 0 0 0 1 −1.5 0 0 3 4 1 3 −1.25 1 0 2 1 0 0 −1 1 2 8 4 0 3−0.75 1 5 6 11 1 0 −0.5 2 6 6 10 1 1 −0.25 4 8 6 3 2 1 0 9 5 8 9 1 00.25 8 6 5 3 2 0 0.5 3 3 6 1 0 0 0.75 4 3 1 1 0 0 1 3 0 2 0 0 0 More 0 01 0 1 0 Gray Matter distribution −2 0 0 1 4 0 1 −1.75 0 1 2 1 0 1 −1.5 11 3 6 0 0 −1.25 2 2 4 6 0 0 −1 2 3 8 4 1 2 −0.75 1 5 6 5 0 1 −0.5 1 6 46 1 2 −0.25 5 6 7 3 2 0 0 4 5 4 3 2 2 0.25 6 3 6 6 2 1 0.5 5 2 2 1 0 00.75 3 1 2 2 0 0 1 3 2 2 0 0 0 More 3 1 3 1 1 0

TABLE 48 Effect of Age on White and Gray Matter Scores in Males WhiteMatter Gray Matter Score Score Mean SEM Mean SEM Cohort Age Ctl, 30-39,Male −0.25 0.29 −0.92 0.28 Age Ctl, 40-49, Male −0.48 0.10 −1.28 0.14Age Ctl, 50-59, Male −0.47 0.08 −0.90 0.11 Age Ctl, 60-69, Male −0.530.10 −0.84 0.14 Age Ctl, 70+_Male −0.43 0.09 −0.78 0.14 Comparison 50-59vs. 40-49, Male Delta 0.01 Delta 0.38 ttest 9.4E−01 ttest 4.2E−02 60-69vs. 40-49, Male Delta −0.05 Delta 0.45 ttest 7.4E−01 ttest 3.2E−02 70+vs. 40-49, Male Delta 0.05 Delta 0.50 ttest 7.2E−01 ttest 1.4E−02

TABLE 49 Effect of Age on White and Gray Matter Scores in Females WhiteMatter Gray Matter Score Score Mean SEM Mean SEM Cohort Age Ctl, 30-39,Female −0.36 0.18 −0.56 0.27 Age Ctl, 40-49, Female −0.33 0.10 −0.610.13 Age Ctl, 50-59, Female −0.55 0.06 −0.71 0.09 Age Ctl, 60-69, Female−0.62 0.09 −0.90 0.12 Age Ctl, 70+_Female −0.47 0.11 −0.58 0.14Comparison 50-59 vs. 40-49, Female Delta −0.22 Delta −0.10 ttest 6.9E−02ttest 5.4E−01 60-69 vs. 40-49, Female Delta −0.29 Delta −0.30 ttest3.6E−02 ttest 1.1E−01 70+ vs. 40-49, Female Delta −0.14 Delta 0.03 ttest3.7E−01 ttest 8.8E−01

TABLE 50 Risk prediction in Males Stats CN Low Moderate Severe PM Ctl PMSDAT White Matter White Matter White Matter White Matter White MatterWhite Matter Score Score Score Score Score Score Control Control ControlControl Control Control Normalized Normalized Normalized NormalizedNormalized Normalized Total N 32 40 58 18 10 10 Total L 27 30 38 8 8 3Total H 5 10 20 10 2 7 L % 84.4 75.0 65.5 44.4 80.0 30.0 Gray MatterGray Matter Gray Matter Gray Matter Gray Matter Gray Matter Score ScoreScore Score Score Score Control Control Control Control Control ControlNormalized Normalized Normalized Normalized Normalized Normalized TotalN 32 40 58 18 10 10 Total L 23 19 26 3 8 1 Total H 9 21 32 15 2 9 L %71.9 47.5 44.8 16.7 80.0 10.0 Low risk LL 22 19 25 3 7 1 % LL 68.8 47.543.1 16.7 70.0 10.0 Intermediate risk IM 6 11 14 5 2 2 % IM 18.8 27.524.1 27.8 20.0 20.0 High risk HH 4 10 19 10 1 7 % HH 12.5 25.0 32.8 55.610.0 70.0

TABLE 51 Risk Prediction in Females Stats CN Low Moderate Severe PM CtlPM SDAT White Matter White Matter White Matter White Matter White MatterWhite Matter Score Score Score Score Score Score Control Control ControlControl Control Control Normalized Normalized Normalized NormalizedNormalized Normalized Total N 36 38 54 48 9 10 Total L 31 25 29 17 6 1Total H 5 13 25 31 3 9 L % 86.1 65.8 53.7 35.4 66.7 10.0 Gray MatterGray Matter Gray Matter Gray Matter Gray Matter Gray Matter Score ScoreScore Score Score Score Control Control Control Control Control ControlNormalized Normalized Normalized Normalized Normalized Normalized TotalN 36 38 54 48 9 10 Total L 29 20 26 16 7 3 Total H 7 18 28 32 2 7 L %80.6 52.6 48.1 33.3 77.8 30.0 Low risk LL 29 17 24 11 5 0 % LL 80.6 44.744.4 22.9 55.6 0.0 Intermediate risk IM 2 11 7 11 3 4 % IM 5.6 28.9 13.022.9 33.3 40.0 High risk HH 5 10 23 26 1 6 % HH 13.9 26.3 42.6 54.2 11.160.0

TABLE 52 Summary of key ratio and p-value statistics for EtnPls16:0/22:6 (M19) to PtdEt 16:0/18:0 (M01) serum ratio for males andfemales combined. Comparison Ratio T-test 60-69 to 50-59 0.75 1.2E−0270-95 to 50-59 0.95 6.4E−01 CN to 50-59 1.07 4.8E−01 SDAT to 50-59 0.704.7E−07 70-95 to 60-69 1.26 6.9E−02 CN to 60-69 1.42 3.8E−04 SDAT to70-95 0.74 1.3E−04 SDAT to CN 0.65 7.6E−11 ADAS 5-19 to CN 0.74 3.0E−04ADAS 20-39 to CN 0.66 1.3E−07 ADAS 40-70 to CN 0.53 3.9E−11 ADAS 20-39to ADAS 5-19 0.90 1.6E−01 ADAS 40-70 to ADAS 5-19 0.72 3.4E−04 ADAS40-70 to ADAS 20-39 0.80 1.0E−02 Post-Mortem SDAT to Control* 0.554.7E−03 *ratio and p-value of EtnPls 16:0/22:6 alone.

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What is claimed:
 1. A method for diagnosing a patient's Alzheimer'sdisease health state or change in health state, or for diagnosingAlzheimer's disease, dementia, cognitive impairment, the risk ofAlzheimer's disease, the risk of cognitive impairment, or the risk ofdementia in a patient, the method comprising the steps of: a) analyzinga blood sample from said patient by high resolution mass spectrometry toobtain accurate mass intensity data; b) comparing the accurate massintensity data to corresponding data obtained from one or more than onereference sample to identify an increase or decrease in accurate massintensity; and c) using said increase or decrease in accurate massintensity for diagnosing said patient's Alzheimer's disease health stateor change in health state, or for diagnosing Alzheimer's disease,dementia, cognitive impairment, the risk of Alzheimer's disease, therisk of cognitive impairment, or the risk of dementia in said patient,wherein the accurate mass intensity is measured, in Daltons, at orsubstantially equivalent to a hydrogen and electron adjusted accuratemass, or neutral accurate mass, selected from the group consisting of:161.1053, 174.1409, 183.0661, 183.0662, 186.1408, 188.1566, 190.0330,195.8577, 197.8549, 200.1564, 202.0453, 202.1720, 203.1157, 204.1876,205.8865, 206.0789, 207.0822, 214.1720, 215.9153, 216.1877, 218.0192,220.0798, 228.1877, 231.8893, 232.2188, 242.2030, 243.0718, 244.0559,244.2187, 246.1465, 246.1472, 250.0366, 252.2095, 253.8163, 253.8164,255.8135, 257.8105, 260.2135, 262.0221, 262.0777, 262.2293, 264.2452,265.8423, 270.2343, 275.8712, 276.2095, 278.2254, 278.2255, 282.2573,283.2602, 283.9028, 287.2824, 294.1443, 294.2207, 295.2239, 295.2287,302.2255, 303.1079, 304.2402, 304.2410, 305.2438, 306.2569, 310.2883,311.7750, 312.2312, 312.2663, 313.1153, 313.2690, 313.7721, 313.7722,315.7693, 320.2356, 320.2357, 321.8039, 328.2402, 336.2662, 338.2821,339.2850, 340.2976, 341.8614, 350.2423, 351.8903, 365.3159, 369.7338,371.3542, 371.7311, 373.7280, 380.3096, 382.1082, 382.1083, 383.3283,383.3284, 392.2938, 399.8196, 401.8166, 409.0208, 409.8484, 411.3211,411.3212, 419.8773, 429.3749, 429.6888, 429.6890, 430.3818, 431.3860,431.9570, 432.1532, 432.3254, 440.3532, 444.2717, 446.3403, 458.2404,458.2405, 467.8070, 469.8042, 474.3706, 478.2540, 478.3664, 481.3172,482.3215, 484.3041, 484.3794, 485.6503, 487.6482, 488.3873, 489.6451,490.3641, 492.3816, 493.3850, 494.4342, 494.4343, 495.3320, 496.3355,501.3212, 504.3814, 505.3229, 506.2851, 506.3213, 506.3214, 507.3316,507.7055, 508.3356, 509.3493, 509.3494, 512.3356, 512.4082, 513.4122,514.6879, 517.3136, 517.3140, 517.3722, 518.3174, 518.4345, 518.4346,519.3321, 520.3353, 520.3354, 520.4499, 520.4500, 521.3474, 521.3477,521.4522, 522.3511, 522.4634, 522.4635, 523.3634, 523.3635, 523.4678,523.4679, 523.4680, 529.3166, 529.3167, 530.3213, 530.3471, 530.3820,530.3821, 535.7944, 536.4794, 538.4224, 541.3139, 541.3430, 541.3432,541.3433, 541.3435, 541.4425, 542.3173, 542.3461, 543.3296, 544.3970,544.3971, 544.4479, 544.4481, 545.3453, 545.6062, 546.3485, 546.3486,547.6031, 548.4815, 549.4844, 549.4845, 549.6005, 550.4956, 550.4957,551.4985, 551.4986, 552.5021, 552.5022, 555.3098, 555.3101, 555.3102,558.4667, 559.4690, 559.4700, 562.4600, 564.5134, 565.3393, 565.3394,565.3396, 565.4104, 566.3434, 566.4130, 567.3545, 567.3547, 568.3573,568.3574, 569.3685, 569.3687, 569.3690, 569.4784, 570.3724, 570.3725,572.4468, 572.4816, 573.4852, 574.4597, 574.4637, 575.2728, 575.4632,575.4990, 576.4770, 576.5097, 576.5098, 577.5136, 577.5137, 578.1574,578.5277, 579.5325, 580.5350, 580.5351, 580.5352, 581.3344, 581.3345,582.2473, 582.6334, 584.2646, 585.2664, 585.2673, 588.4731, 589.3403,589.3404, 590.3430, 590.3431, 591.3542, 592.3571, 593.4743, 594.4878,595.4932, 596.4791, 597.4840, 598.4965, 598.5124, 599.4936, 600.5127,601.5164, 602.5282, 603.5317, 604.5432, 604.5433, 605.5456, 605.5457,606.4869, 607.4919, 610.3686, 613.3405, 614.4914, 615.3539, 615.4798,615.4938, 616.4673, 616.5052, 617.5089, 618.4829, 622.2539, 624.5131,625.5075, 625.5076, 625.5161, 626.5104, 626.5108, 626.5277, 626.5278,627.5201, 627.5302, 627.5304, 628.5237, 628.5421, 629.5449, 630.4861,630.5586, 630.5587, 631.4902, 631.5619, 631.6280, 632.5032, 632.5035,632.5762, 633.5078, 638.2465, 638.5149, 640.2637, 640.5294, 641.2661,646.4837, 651.5221, 653.5360, 653.5361, 656.2587, 661.6233, 662.5175,663.4870, 663.5216, 664.5322, 664.5323, 665.5010, 665.5354, 666.5455,666.5456, 667.5474, 667.5475, 670.5689, 670.5690, 671.5722, 671.5723,671.5726, 675.6357, 675.6377, 678.5477, 680.5623, 684.5489, 684.5491,685.2600, 685.2601, 685.5538, 686.4877, 686.4880, 691.1957, 691.5960,694.6161, 699.5198, 699.5199, 699.5200, 699.5203, 700.5520, 701.5359,702.5677, 703.5516, 703.5709, 705.6093, 709.2594, 709.2595, 711.2577,715.5167, 715.5169, 717.5308, 719.5465, 719.6231, 719.6233, 720.6258,720.6438, 721.6390, 722.4789, 722.5334, 722.5335, 722.6424, 723.5194,723.5195, 723.5197, 723.5203, 724.5256, 724.5257, 724.5258, 724.5493,725.5359, 725.5385, 725.5527, 726.5459, 726.5461, 726.5462, 727.5516,727.5567, 727.5568, 727.5569, 728.5626, 728.5627, 728.5629, 729.5672,730.6493, 731.4914, 731.4916, 731.4917, 731.5463, 731.5464, 731.5829,731.6530, 732.4937, 732.4938, 732.4939, 732.4940, 733.6426, 738.5445,738.5448, 739.5141, 739.5142, 739.5143, 739.5164, 741.5305, 741.5307,741.5319, 742.2972, 743.5464, 743.5466, 743.5467, 743.5468, 743.5469,743.5470, 743.5471, 744.4954, 744.4956, 744.5500, 744.5535, 744.5536,745.5621, 745.5656, 745.5658, 746.5119, 746.5570, 746.5714, 746.5717,746.5719, 746.6210, 747.5201, 747.5203, 747.5244, 747.5245, 747.5778,748.5286, 748.5287, 748.5721, 748.5735, 749.5359, 749.5365, 749.5367,749.5406, 749.5407, 749.5408, 749.5760, 749.5761, 749.5777, 750.5402,750.5403, 750.5438, 750.5440, 751.5516, 751.5529, 751.5553, 751.5555,751.5556, 752.5564, 752.5581, 752.5583, 753.5672, 753.5672, 755.4860,755.4861, 755.5467, 755.5468, 757.4989, 757.4991, 757.4993, 757.5014,757.5016, 757.5625, 757.5626, 758.5092, 758.5094, 758.5655, 758.5656,759.5162, 759.5163, 759.5164, 759.5165, 759.5779, 760.5215, 760.5216,760.5217, 760.5811, 761.5843, 761.5846, 763.5157, 765.5313, 765.5334,765.5335, 767.5469, 767.5470, 767.5494, 767.5495, 768.5503, 768.5504,768.5538, 768.5539, 769.5654, 769.5655, 769.5656, 771.5813, 771.5814,771.5815, 772.5278, 772.5280, 772.5860, 772.5862, 772.5863, 773.5347,773.5368, 773.5370, 774.0313, 774.0316, 775.5516, 775.5527, 775.5528,775.5530, 775.5533, 775.5535, 777.4426, 777.5287, 777.5288, 777.5530,777.5531, 777.5672, 779.4393, 779.5441, 779.5442, 779.5444, 779.5829,780.5474, 780.5475, 781.4354, 781.5607, 781.5616, 781.5617, 781.5620,781.5985, 782.5084, 782.5085, 782.5086, 782.5647, 782.5649, 782.5650,782.5653, 783.4315, 783.5147, 783.5148, 783.5671, 783.5672, 783.5778,784.5235, 784.5237, 784.5239, 784.5810, 784.5811, 785.4288, 785.5933,786.5415, 786.5416, 786.5967, 787.5464, 787.5465, 787.5728, 787.5729,789.5892, 789.5893, 793.5386, 793.5387, 793.5679, 793.5681, 794.5421,794.5422, 795.5181, 795.5182, 795.5183, 795.5550, 795.5551, 795.5837,795.5838, 795.5839, 796.5290, 796.5292, 796.5874, 796.5876, 798.6742,801.5550, 802.7056, 803.5445, 803.5677, 803.5678, 803.5680, 803.5681,804.5475, 804.5476, 804.5713, 804.5718, 804.7227, 804.7230, 805.5604,805.5605, 805.5831, 805.5832, 805.5839, 806.5637, 806.5638, 806.5639,806.5873, 807.5757, 807.5758, 807.5768, 807.5899, 807.5900, 808.5792,808.5803, 809.5936, 809.5937, 810.5399, 810.5400, 810.5401, 810.5969,810.5971, 811.5732, 811.6096, 812.5761, 812.5762, 813.5885, 814.5917,814.5919, 817.5375, 817.5376, 817.5377, 817.5378, 817.5838, 817.5840,819.5551, 819.5552, 819.5553, 819.5641, 819.5642, 820.5677, 820.5679,820.5680, 821.5711, 821.5712, 821.5713, 824.6892, 825.5544, 825.6926,825.6927, 826.5581, 826.7048, 826.7069, 826.7070, 827.5448, 827.5699,827.5700, 827.5701, 827.7083, 827.7086, 828.5736, 828.5737, 828.5742,828.5743, 828.7202, 828.7207, 829.5604, 829.5856, 829.5859, 829.7239,829.7242, 829.7244, 829.7246, 830.5894, 830.7352, 830.7355, 830.7362,830.7363, 831.5759, 831.5760, 831.5995, 831.5997, 831.5998, 831.6000,831.6001, 831.7387, 831.7408, 832.5791, 832.5792, 832.6024, 832.6026,832.6027, 832.6036, 832.6037, 832.7492, 832.7495, 832.7521, 832.7522,832.7523, 833.5931, 833.5932, 833.7551, 833.7558, 833.7570, 833.7571,835.6998, 835.7001, 835.7006, 836.7076, 837.5027, 837.5881, 837.7180,838.7226, 838.7232, 839.6031, 840.6063, 842.7387, 847.5316, 847.5953,847.5954, 851.5698, 852.5738, 852.7250, 853.5854, 853.5855, 853.5862,853.7296, 854.5902, 855.6015, 855.6016, 855.6023, 855.6025, 855.7417,856.6061, 856.6063, 856.6720, 856.7481, 857.6186, 857.7531, 858.6211,858.6212, 858.6860, 858.6861, 858.7632, 858.7663, 859.7694, 859.7695,859.7696, 859.7706, 860.7752, 860.7753, 860.7756, 861.7801, 861.7806,863.6876, 863.7336, 864.7380, 864.7596, 865.7486, 865.7487, 866.7532,866.7533, 867.7579, 867.7581, 870.7307, 871.5527, 871.5528, 871.5529,871.5934, 871.5935, 874.7645, 878.7400, 879.5999, 879.7454, 880.6035,880.7516, 880.7566, 881.7558, 881.7616, 882.7660, 882.7723, 883.7705,883.7765, 884.7801, 884.7877, 885.7854, 885.7922, 886.5582, 886.5584,886.7916, 886.7917, 886.8012, 887.7352, 887.7999, 887.8001, 887.8005,888.7394, 889.7492, 890.7535, 894.7810, 908.7832, 909.7882, 910.7968,911.8032, 913.7513, 914.7583, 915.5191, 915.5192, 915.7681, 916.7743,921.8142, 921.8145, 921.8153, 928.7505, 931.7695, 942.7879, 1018.9420,1019.3838, 1019.3840, 1085.3294, 1098.9739, 1098.9740, 1225.0920,1226.0968, 1226.0970, 1227.1070, 1228.1110, 1228.1111 and combinationsthereof.
 2. The method of claim 1, wherein the accurate mass intensitydata is obtained using a Fourier transform ion cyclotron resonance, timeof flight, orbitrap, quadrupole or triple quadrupole mass spectrometer.3. The method of claim 1, wherein the blood sample is a blood serumsample.
 4. The method of claim 1, wherein the accurate mass intensitiesrepresent ionized metabolites.
 5. The method of claim 1, wherein aliquid/liquid extraction is performed on the blood sample wherebynon-polar metabolites are dissolved in an organic solvent and polarmetabolites are dissolved in an aqueous solvent.
 6. The method of claim5, wherein the accurate mass intensities are obtained from theionization of the extracted samples using an ionization method selectedfrom the group consisting of: positive electrospray ionization, negativeelectrospray ionization, positive atmospheric pressure chemicalionization, negative atmospheric pressure chemical ionization, andcombinations thereof.
 7. The method of claim 1, wherein the accuratemass intensity data is obtained using a Fourier transform ion cyclotronresonance mass spectrometer.
 8. The method of claim 1, wherein said oneor more than one reference sample is: a) one or more than one referencesample obtained from a non-demented control individual; b) one or morethan one reference sample obtained from a demented subject; c) one ormore than one reference sample obtained from a subject pathologicallydiagnosed as having Alzheimer's Disease; d) one or more than onereference sample obtained from a subject clinically diagnosed as havingAlzheimer's Disease; e) one or more than one reference sample obtainedfrom a patient with cognitive impairment as measured by Alzheimer'sDisease Assessment Scale-cognitive subset (ADAS-cog); f) one or morethan one reference sample obtained from a patient with cognitiveimpairment as measured by Folstein's Mini-Mental State Exam (MMSE); org) any combination of a) to f) above.
 9. The method of claim 1, whereinthe method is for diagnosing: (a) the presence of or risk of Alzheimer'sdisease (AD); (b) the presence of or risk of AD pathology; (c) thepresence of or risk of dementia; (d) the presence of or risk ofcognitive impairment; or (e) a combination of (a) to (d) above.
 10. Themethod of claim 1, further comprising analyzing a blood sample from saidpatient by mass spectrometry to obtain accurate mass intensity data forone or more than one internal control metabolite; and calculating aratio for each of the accurate mass intensities obtained in step (a) tothe accurate mass intensities obtained for the one or more than oneinternal control metabolite; wherein the comparing step (b) comprisescomparing each ratio to one or more corresponding ratios obtained forone or more than one reference sample.
 11. The method of claim 1,wherein a decrease in accurate mass intensity is identified in thecomparing step (b).
 12. The method of claim 1, wherein the hydrogen andelectron adjusted accurate mass, or neutral accurate mass, is selectedfrom the group consisting of: 699.5198, 723.5195, 723.5197, 751.5555,541.3432, 569.3687, 803.568, 886.5582, 565.3394, 569.369, 801.555,857.6186, 207.0822, 275.8712, 371.7311, 373.728, 432.1532, 485.5603,487.6482, 562.46, 622.2539, 640.2637, 730.6493, 742.2972, 701.53591,699.52026, 723.52026, 747.52026, 729.56721, 727.55156, 779.58286,775.55156, and combinations thereof.
 13. A method for diagnosing apatient's Alzheimer's disease health state or change in health state, orfor diagnosing Alzheimer's disease, dementia, cognitive impairment, therisk of Alzheimer's disease, the risk of cognitive impairment, or therisk of dementia in a patient, the method comprising the steps of: a)analyzing a blood sample from said patient to obtain quantifying datafor one or more than one metabolite marker; b) comparing the quantifyingdata for said one or more than one metabolite marker to correspondingdata obtained for one or more than one reference sample to identify anincrease or decrease in the level of said one or more than onemetabolite marker in said blood sample; and c) using said increase ordecrease in the level of said one or more than one metabolite marker insaid blood sample for diagnosing said patient's Alzheimer's diseasehealth state or change in health state, or for diagnosing Alzheimer'sdisease, dementia, cognitive impairment, the risk of Alzheimer'sdisease, the risk of cognitive impairment, or the risk of dementia insaid patient, wherein the one or more than one metabolite markercomprises one or more molecule selected from the group consisting of:phosphatidylethanolamine (PtdEt), plasmenylethanolamine,plasmanylethanolamine, and combinations thereof, wherein the PtdEt isselected from the group consisting of PtdEt 16:0/18:0, PtdEt 16:0/18:1,PtdEt 18:0/18:0, PtdEt 18:0/18:1, and combinations thereof.
 14. Themethod of claim 13, wherein the-plasmanylethanolamine is selected fromthe group consisting of: plasmanyl 16:0/18:1, plasmanyl 16:0/18:2,plasmanyl 16:0/20:4, plasmanyl 16:0/22:4, plasmanyl 16:0/22:6, plasmanyl18:0/18:1, plasmanyl 18:0/18:2, plasmanyl 18:0/20:4, plasmanyl18:0/22:4, plasmanyl 18:0/22:6, and combinations thereof, and theplasmenylethanolamine is selected from the group consisting of:plasmenyl 16:0/18:1, plasmenyl 16:0/18:2, plasmenyl 16:0/20:4, plasmenyl16:0/22:4, plasmenyl 16:0/22:6, plasmenyl 18:0/18:1, plasmenyl18:0/18:2, plasmenyl 18:0/20:4, plasmenyl 18:0/22:4, plasmenyl18:0/22:6, and combinations thereof.
 15. The method of claim 13, whereinthe quantifying data is obtained using a Fourier transform ion cyclotronresonance, time of flight, orbitrap, quadrupole or triple quadrupolemass spectrometer.
 16. The method of claim 15, wherein the massspectrometer is equipped with a chromatographic system.
 17. The methodof claim 13, wherein the blood sample is a blood serum sample.
 18. Themethod of claim 13, wherein a liquid/liquid extraction is performed onthe blood sample whereby non-polar metabolites are dissolved in anorganic solvent and polar metabolites are dissolved in an aqueoussolvent.
 19. The method of claim 18, wherein the extracted samples areanalyzed by positive or negative electrospray ionization or positive ornegative atmospheric pressure chemical ionization.
 20. The method ofclaim 18, wherein the extracted samples are analyzed by MS/MStransition.
 21. The method of claim 18, wherein the extracted samplesare analyzed by extracted ion current (EIC) chromatography and MS/MStransition.
 22. The method of claim 13, wherein said one or more thanone reference sample is a) one or more than one reference sampleobtained from a non-demented control individual; b) one or more than onereference sample obtained from a demented subject; c) one or more thanone reference sample obtained from a subject pathologically diagnosed ashaving Alzheimer's Disease; d) one or more than one reference sampleobtained from a subject clinically diagnosed as having Alzheimer'sDisease; e) one or more than one reference sample obtained from apatient with cognitive impairment as measured by Alzheimer's DiseaseAssessment Scale-cognitive subset (ADAS-cog); f) one or more than onereference sample obtained from a patient with cognitive impairment asmeasured by Folstein's Mini-Mental State Exam (MMSE); or g) anycombination of a) to f) above.
 23. The method of claim 13, furthercomprising: analyzing a blood sample from said patient to obtainquantifying data for one or more than one internal control metabolite;and obtaining a ratio for each of the levels of said one or more thanone metabolite marker to the level obtained for the one or more than oneinternal control metabolite; wherein the comparing step (b) comprisescomparing each ratio to one or more corresponding ratios obtained forthe one or more than one reference sample.
 24. The method of claim 13,wherein a decrease in the level of said one or more than one metabolitemarker in said blood sample is identified in the comparing step (b). 25.The method of claim 14, wherein the extracted samples are analyzed byMS/MS transitions, and the MS/MS transitions for thephosphatidylethanolamines are 718.0/255.0, 716.0/255.0, 746.0/283.0 and744.0/283.0, respectively; the MS/MS transitions for theplasmanylethanolamines are 702.0/281.0, 700.0/279.0, 724.0/303.0,752.0/331.0, 748.0/327.0, 730.0/281.0, 728.0/279.0, 752.0/303.0,780.0/331.0 and 776.0/327.0, respectively; and the MS/MS transitions forthe plasmenylethanolamines are 700.0/281.0, 698.0/279.0, 722.0/303.0,750.0/331.0, 746.0/327.0, 728.0/281.0, 726.0/279.0, 750.6/303.2,778.0/331.0 and 774.0/327.0, respectively.
 26. A method for evaluatingthe efficacy of a therapy for treating Alzheimer's disease, dementia, orcognitive impairment in a patient, comprising the steps of: a) analyzinga blood sample from said patient by high resolution mass spectrometry toobtain accurate mass intensity data; b) comparing the accurate massintensity data to corresponding data obtained from one or more than onereference sample to identify an increase or decrease in accurate massintensity; and c) using said increase or decrease in accurate massintensity to determine whether the therapy is improving the biochemicalstate of the patient, wherein the accurate mass intensity is measured,in Daltons, at or substantially equivalent to a hydrogen and electronadjusted accurate mass, or neutral accurate mass, selected from thegroup consisting of: 161.1053, 174.1409, 183.0661, 183.0662, 186.1408,188.1566, 190.0330, 195.8577, 197.8549, 200.1564, 202.0453, 202.1720,203.1157, 204.1876, 205.8865, 206.0789, 207.0822, 214.1720, 215.9153,216.1877, 218.0192, 220.0798, 228.1877, 231.8893, 232.2188, 242.2030,243.0718, 244.0559, 244.2187, 246.1465, 246.1472, 250.0366, 252.2095,253.8163, 253.8164, 255.8135, 257.8105, 260.2135, 262.0221, 262.0777,262.2293, 264.2452, 265.8423, 270.2343, 275.8712, 276.2095, 278.2254,278.2255, 282.2573, 283.2602, 283.9028, 287.2824, 294.1443, 294.2207,295.2239, 295.2287, 302.2255, 303.1079, 304.2402, 304.2410, 305.2438,306.2569, 310.2883, 311.7750, 312.2312, 312.2663, 313.1153, 313.2690,313.7721, 313.7722, 315.7693, 320.2356, 320.2357, 321.8039, 328.2402,336.2662, 338.2821, 339.2850, 340.2976, 341.8614, 350.2423, 351.8903,365.3159, 369.7338, 371.3542, 371.7311, 373.7280, 380.3096, 382.1082,382.1083, 383.3283, 383.3284, 392.2938, 399.8196, 401.8166, 409.0208,409.8484, 411.3211, 411.3212, 419.8773, 429.3749, 429.6888, 429.6890,430.3818, 431.3860, 431.9570, 432.1532, 432.3254, 440.3532, 444.2717,446.3403, 458.2404, 458.2405, 467.8070, 469.8042, 474.3706, 478.2540,478.3664, 481.3172, 482.3215, 484.3041, 484.3794, 485.6503, 487.6482,488.3873, 489.6451, 490.3641, 492.3816, 493.3850, 494.4342, 494.4343,495.3320, 496.3355, 501.3212, 504.3814, 505.3229, 506.2851, 506.3213,506.3214, 507.3316, 507.7055, 508.3356, 509.3493, 509.3494, 512.3356,512.4082, 513.4122, 514.6879, 517.3136, 517.3140, 517.3722, 518.3174,518.4345, 518.4346, 519.3321, 520.3353, 520.3354, 520.4499, 520.4500,521.3474, 521.3477, 521.4522, 522.3511, 522.4634, 522.4635, 523.3634,523.3635, 523.4678, 523.4679, 523.4680, 529.3166, 529.3167, 530.3213,530.3471, 530.3820, 530.3821, 535.7944, 536.4794, 538.4224, 541.3139,541.3430, 541.3432, 541.3433, 541.3435, 541.4425, 542.3173, 542.3461,543.3296, 544.3970, 544.3971, 544.4479, 544.4481, 545.3453, 545.6062,546.3485, 546.3486, 547.6031, 548.4815, 549.4844, 549.4845, 549.6005,550.4956, 550.4957, 551.4985, 551.4986, 552.5021, 552.5022, 555.3098,555.3101, 555.3102, 558.4667, 559.4690, 559.4700, 562.4600, 564.5134,565.3393, 565.3394, 565.3396, 565.4104, 566.3434, 566.4130, 567.3545,567.3547, 568.3573, 568.3574, 569.3685, 569.3687, 569.3690, 569.4784,570.3724, 570.3725, 572.4468, 572.4816, 573.4852, 574.4597, 574.4637,575.2728, 575.4632, 575.4990, 576.4770, 576.5097, 576.5098, 577.5136,577.5137, 578.1574, 578.5277, 579.5325, 580.5350, 580.5351, 580.5352,581.3344, 581.3345, 582.2473, 582.6334, 584.2646, 585.2664, 585.2673,588.4731, 589.3403, 589.3404, 590.3430, 590.3431, 591.3542, 592.3571,593.4743, 594.4878, 595.4932, 596.4791, 597.4840, 598.4965, 598.5124,599.4936, 600.5127, 601.5164, 602.5282, 603.5317, 604.5432, 604.5433,605.5456, 605.5457, 606.4869, 607.4919, 610.3686, 613.3405, 614.4914,615.3539, 615.4798, 615.4938, 616.4673, 616.5052, 617.5089, 618.4829,622.2539, 624.5131, 625.5075, 625.5076, 625.5161, 626.5104, 626.5108,626.5277, 626.5278, 627.5201, 627.5302, 627.5304, 628.5237, 628.5421,629.5449, 630.4861, 630.5586, 630.5587, 631.4902, 631.5619, 631.6280,632.5032, 632.5035, 632.5762, 633.5078, 638.2465, 638.5149, 640.2637,640.5294, 641.2661, 646.4837, 651.5221, 653.5360, 653.5361, 656.2587,661.6233, 662.5175, 663.4870, 663.5216, 664.5322, 664.5323, 665.5010,665.5354, 666.5455, 666.5456, 667.5474, 667.5475, 670.5689, 670.5690,671.5722, 671.5723, 671.5726, 675.6357, 675.6377, 678.5477, 680.5623,684.5489, 684.5491, 685.2600, 685.2601, 685.5538, 686.4877, 686.4880,691.1957, 691.5960, 694.6161, 699.5198, 699.5199, 699.5200, 699.5203,700.5520, 701.5359, 702.5677, 703.5516, 703.5709, 705.6093, 709.2594,709.2595, 711.2577, 715.5167, 715.5169, 717.5308, 719.5465, 719.6231,719.6233, 720.6258, 720.6438, 721.6390, 722.4789, 722.5334, 722.5335,722.6424, 723.5194, 723.5195, 723.5197, 723.5203, 724.5256, 724.5257,724.5258, 724.5493, 725.5359, 725.5385, 725.5527, 726.5459, 726.5461,726.5462, 727.5516, 727.5567, 727.5568, 727.5569, 728.5626, 728.5627,728.5629, 729.5672, 730.6493, 731.4914, 731.4916, 731.4917, 731.5463,731.5464, 731.5829, 731.6530, 732.4937, 732.4938, 732.4939, 732.4940,733.6426, 738.5445, 738.5448, 739.5141, 739.5142, 739.5143, 739.5164,741.5305, 741.5307, 741.5319, 742.2972, 743.5464, 743.5466, 743.5467,743.5468, 743.5469, 743.5470, 743.5471, 744.4954, 744.4956, 744.5500,744.5535, 744.5536, 745.5621, 745.5656, 745.5658, 746.5119, 746.5570,746.5714, 746.5717, 746.5719, 746.6210, 747.5201, 747.5203, 747.5244,747.5245, 747.5778, 748.5286, 748.5287, 748.5721, 748.5735, 749.5359,749.5365, 749.5367, 749.5406, 749.5407, 749.5408, 749.5760, 749.5761,749.5777, 750.5402, 750.5403, 750.5438, 750.5440, 751.5516, 751.5529,751.5553, 751.5555, 751.5556, 752.5564, 752.5581, 752.5583, 753.5672,753.5672, 755.4860, 755.4861, 755.5467, 755.5468, 757.4989, 757.4991,757.4993, 757.5014, 757.5016, 757.5625, 757.5626, 758.5092, 758.5094,758.5655, 758.5656, 759.5162, 759.5163, 759.5164, 759.5165, 759.5779,760.5215, 760.5216, 760.5217, 760.5811, 761.5843, 761.5846, 763.5157,765.5313, 765.5334, 765.5335, 767.5469, 767.5470, 767.5494, 767.5495,768.5503, 768.5504, 768.5538, 768.5539, 769.5654, 769.5655, 769.5656,771.5813, 771.5814, 771.5815, 772.5278, 772.5280, 772.5860, 772.5862,772.5863, 773.5347, 773.5368, 773.5370, 774.0313, 774.0316, 775.5516,775.5527, 775.5528, 775.5530, 775.5533, 775.5535, 777.4426, 777.5287,777.5288, 777.5530, 777.5531, 777.5672, 779.4393, 779.5441, 779.5442,779.5444, 779.5829, 780.5474, 780.5475, 781.4354, 781.5607, 781.5616,781.5617, 781.5620, 781.5985, 782.5084, 782.5085, 782.5086, 782.5647,782.5649, 782.5650, 782.5653, 783.4315, 783.5147, 783.5148, 783.5671,783.5672, 783.5778, 784.5235, 784.5237, 784.5239, 784.5810, 784.5811,785.4288, 785.5933, 786.5415, 786.5416, 786.5967, 787.5464, 787.5465,787.5728, 787.5729, 789.5892, 789.5893, 791.5386, 793.5387, 791.5679,793.5681, 794.5421, 794.5422, 795.5181, 795.5182, 795.5183, 795.5550,795.5551, 795.5837, 795.5838, 795.5839, 796.5290, 796.5292, 796.5874,796.5876, 798.6742, 801.5550, 802.7056, 803.5445, 803.5677, 803.5678,803.5680, 803.5681, 804.5475, 804.5476, 804.5713, 804.5718, 804.7227,804.7230, 805.5604, 805.5605, 805.5831, 805.5832, 805.5839, 806.5637,806.5638, 806.5639, 806.5873, 807.5757, 807.5758, 807.5768, 807.5899,807.5900, 808.5792, 808.5803, 809.5936, 809.5937, 810.5399, 810.5400,810.5401, 810.5969, 810.5971, 811.5732, 811.6096, 812.5761, 812.5762,813.5885, 814.5917, 814.5919, 817.5375, 817.5376, 817.5377, 817.5378,817.5838, 817.5840, 819.5551, 819.5552, 819.5553, 819.5641, 819.5642,820.5677, 820.5679, 820.5680, 821.5711, 821.5712, 821.5713, 824.6892,825.5544, 825.6926, 825.6927, 826.5581, 826.7048, 826.7069, 826.7070,827.5448, 827.5699, 827.5700, 827.5701, 827.7083, 827.7086, 828.5736,828.5737, 828.5742, 828.5743, 828.7202, 828.7207, 829.5604, 829.5856,829.5859, 829.7239, 829.7242, 829.7244, 829.7246, 830.5894, 830.7352,830.7355, 830.7362, 830.7363, 831.5759, 831.5760, 831.5995, 831.5997,831.5998, 831.6000, 831.6001, 831.7387, 831.7408, 832.5791, 832.5792,832.6024, 832.6026, 832.6027, 832.6036, 832.6037, 832.7492, 832.7495,832.7521, 832.7522, 832.7523, 833.5931, 833.5932, 833.7551, 833.7558,833.7570, 833.7571, 835.6998, 835.7001, 835.7006, 836.7076, 837.5027,837.5881, 837.7180, 838.7226, 838.7232, 839.6031, 840.6063, 842.7387,847.5316, 847.5953, 847.5954, 851.5698, 852.5738, 852.7250, 853.5854,853.5855, 853.5862, 853.7296, 854.5902, 855.6015, 855.6016, 855.6023,855.6025, 855.7417, 856.6061, 856.6063, 856.6720, 856.7481, 857.6186,857.7531, 858.6211, 858.6212, 858.6860, 858.6861, 858.7632, 858.7663,859.7694, 859.7695, 859.7696, 859.7706, 860.7752, 860.7753, 860.7756,861.7801, 861.7806, 863.6876, 863.7336, 864.7380, 864.7596, 865.7486,865.7487, 866.7532, 866.7533, 867.7579, 867.7581, 870.7307, 871.5527,871.5528, 871.5529, 871.5934, 871.5935, 874.7645, 878.7400, 879.5999,879.7454, 880.6035, 880.7516, 880.7566, 881.7558, 881.7616, 882.7660,882.7723, 883.7705, 883.7765, 884.7801, 884.7877, 885.7854, 885.7922,886.5582, 886.5584, 886.7916, 886.7917, 886.8012, 887.7352, 887.7999,887.8001, 887.8005, 888.7394, 889.7492, 890.7535, 894.7810, 908.7832,909.7882, 910.7968, 911.8032, 913.7513, 914.7583, 915.5191, 915.5192,915.7681, 916.7743, 921.8142, 921.8145, 921.8153, 928.3505, 931.7695,942.7879, 1018.9420, 1019.3838, 1019.3840, 1085.3294, 1098.9739,1098.9740, 1225.0920, 1226.0968, 1226.0970, 1227.1070, 1228.1110,1228.1111 and combinations thereof.
 27. The method of claim 26, whereinthe accurate mass intensity data is obtained using a Fourier transformion cyclotron resonance, time of flight, orbitrap, quadrupole or triplequadrupole mass spectrometer.
 28. The method of claim 26, wherein theblood sample is a blood serum sample.
 29. The method of claim 26,wherein the accurate mass intensities represent ionized metabolites. 30.The method of claim 26, wherein a liquid/liquid extraction is performedon the blood sample whereby non-polar metabolites are dissolved in anorganic solvent and polar metabolites are dissolved in an aqueoussolvent.
 31. The method of claim 30, wherein the accurate massintensities are obtained from the ionization of the extracted samplesusing an ionization method selected from the group consisting of:positive electrospray ionization, negative electrospray ionization,positive atmospheric pressure chemical ionization, negative atmosphericpressure chemical ionization, and combinations thereof.
 32. The methodof claim 26, wherein the accurate mass intensity data is obtained usinga Fourier transform ion cyclotron resonance mass spectrometer.
 33. Themethod of claim 26, wherein said one or more than one reference sampleis: a) one or more than one reference sample obtained from anon-demented control individual; b) one or more than one referencesample obtained from a demented subject; c) one or more than onereference sample obtained from a subject pathologically diagnosed ashaving Alzheimer's Disease; d) one or more than one reference sampleobtained from a subject clinically diagnosed as having Alzheimer'sDisease; e) one or more than one reference sample obtained from apatient with cognitive impairment as measured by Alzheimer's DiseaseAssessment Scale-cognitive subset (ADAS-cog); f) one or more than onereference sample obtained from a patient with cognitive impairment asmeasured by Folstein's Mini-Mental State Exam (MMSE); or g) anycombination of a) to above.
 34. The method of claim 26, wherein themethod is for evaluating the efficacy of a therapy for treating: (a)Alzheimer's disease (AD); (b) AD pathology; (c) dementia; (d) cognitiveimpairment; or (e) a combination of (a) to (d) above.
 35. The method ofclaim 26, further comprising analyzing a blood sample from said patientby mass spectrometry to obtain accurate mass intensity data for one ormore than one internal control metabolite; and calculating a ratio foreach of the accurate mass intensities obtained in step (a) to theaccurate mass intensities obtained for the one or more than one internalcontrol metabolite; wherein the comparing step (b) comprises comparingeach ratio to one or more corresponding ratios obtained for one or morethan one reference sample.
 36. The method of claim 26, wherein anincrease in accurate mass intensity is identified in the comparing step(b).
 37. The method of claim 26, wherein the hydrogen and electronadjusted accurate mass, or neutral accurate mass, is selected from thegroup consisting of: 699.5198, 723.5195, 723.5197, 751.5555, 541.3432,569.3687, 803.568, 886.5582, 565.3394, 569.369, 801.555, 857.6186,207.0822, 275.8712, 371.7311, 373.728, 432.1532, 485.5603, 487.6482,562.46, 622.2539, 640.2637, 730.6493, 742.2972, 701.53591, 699.52026,723.52026, 747.52026, 729.56721, 727.55156, 779.58286, 775.55156, andcombinations thereof.
 38. A method for evaluating the efficacy of atherapy for treating Alzheimer's disease, dementia, or cognitiveimpairment in a patient, comprising the steps of: a) analyzing a bloodsample from said patient to obtain quantifying data for one or more thanone metabolite marker; b) comparing the quantifying data for said one ormore than one metabolite marker to corresponding data obtained for oneor more than one reference sample to identify an increase or decrease inthe level of said one or more than one metabolite marker in said bloodsample; and c) using said increase or decrease in the level of said oneor more than one metabolite marker in said blood sample to determinewhether the therapy is improving the biochemical state of the patient,wherein the one or more than one metabolite marker comprises one or moremolecule selected from the group consisting of: phosphatidylethanolamine(PtdEt), plasmenylethanolamine, plasmanylethanolamine, and combinationsthereof, wherein the PtdEt is selected from the group consisting of:PtdEt 16:0/18:0, PtdEt 16:0/18:1, PtdEt 18:0/18:0, PtdEt 18:0/18:1, andcombinations thereof.
 39. The method of claim 38, wherein theplasmanylethanolamine is selected from the group consisting of:plasmanyl 16:0/18:1, plasmanyl 16:0/18:2, plasmanyl 16:0/20:4, plasmanyl16:0/22:4, plasmanyl 16:0/22:6, plasmanyl 18:0/18:1, plasmanyl18:0/18:2, plasmanyl 18:0/20:4, plasmanyl 18:0/22:4, plasmanyl18:0/22:6, and combinations thereof; and, the plasmenylethanolamine isselected from the group consisting of: plasmenyl 16:0/18:1, plasmenyl16:0/18:2, plasmenyl 16:0/20:4, plasmenyl 16:0/22:4, plasmenyl16:0/22:6, plasmenyl 18:0/18:1, plasmenyl 18:0/18:2, plasmenyl18:0/20:4, plasmenyl 18:0/22:4, plasmenyl 18:0/22:6, and combinationsthereof.
 40. The method of claim 38, wherein the quantifying data isobtained using a Fourier transform ion cyclotron resonance, time offlight, orbitrap, quadrupole or triple quadrupole mass spectrometer. 41.The method of claim 40, wherein the mass spectrometer is equipped with achromatographic system.
 42. The method of claim 38, wherein the bloodsample is a blood serum sample.
 43. The method of claim 38, wherein aliquid/liquid extraction is performed on the blood sample wherebynon-polar metabolites are dissolved in an organic solvent and polarmetabolites are dissolved in an aqueous solvent.
 44. The method of claim43, wherein the extracted samples are analyzed by positive or negativeelectrospray ionization or positive or negative atmospheric pressurechemical ionization.
 45. The method of claim 43, wherein the extractedsamples are analyzed by MS/MS transition.
 46. The method of claim 43,wherein the extracted samples are analyzed by extracted ion current(EIC) chromatography and MS/MS transition.
 47. The method of claim 38,wherein said one or more than one reference sample is a) one or morethan one reference sample obtained from a non-demented controlindividual; b) one or more than one reference sample obtained from ademented subject; c) one or more than one reference sample obtained froma subject pathologically diagnosed as having Alzheimer's Disease; d) oneor more than one reference sample obtained from a subject clinicallydiagnosed as having Alzheimer's Disease; e) one or more than onereference sample obtained from a patient with cognitive impairment asmeasured by Alzheimer's Disease Assessment Scale-cognitive subset(ADAS-cog); f) one or more than one reference sample obtained from apatient with cognitive impairment as measured by Folstein's Mini-MentalState Exam (MMSE); or g) any combination of a) to f) above.
 48. Themethod according to claim 38, further comprising: analyzing a bloodsample from said patient to obtain quantifying data for one or more thanone internal control metabolite; and obtaining a ratio for each of thelevels of said one or more than one metabolite marker to the levelobtained for the one or more than one internal control metabolite;wherein the comparing step (b) comprises comparing each ratio to one ormore corresponding ratios obtained for the one or more than onereference sample.
 49. The method of claim 38, wherein an increase in thelevel of said one or more than one metabolite marker in said bloodsample is identified in the comparing step (b).
 50. The method of claim39, wherein the extracted samples are analyzed by MS/MS transition, andthe MS/MS transitions for the phosphatidylethanolamines are 718.0/255.0,716.0/255.0, 746.0/283.0 and 744.0/283.0, respectively; the MS/MStransitions for the plasmanylethanolamines are 702.0/281.0, 700.0/279.0,724.0/303.0, 752.0/331.0, 748.0/327.0, 730.0/281.0, 728.0/279.0,752.0/303.0, 780.0/331.0 and 776.0/327.0, respectively; and the MS/MStransitions for the plasmenylethanolamines are 700.0/281.0, 698.0/279.0,722.0/303.0, 750.0/331.0, 746.0/327.0, 728.0/281.0, 726.0/279.0,750.6/303.2, 778.0/331.0 and 774.0/327.0, respectively.
 51. The methodof claim 26, wherein at least one of the one or more than one referencesample defined in step (b) is a pre-therapy baseline blood sampleobtained from the patient.
 52. The method of claim 1, whereinsubstantially equivalent refers to ±5 ppm of the hydrogen and electronadjusted accurate mass, or neutral accurate mass.
 53. The method ofclaim 1, wherein substantially equivalent refers to ±1 ppm of thehydrogen and electron adjusted accurate mass, or neutral accurate mass.54. The method of claim 26, wherein substantially equivalent refers to±5 ppm of the hydrogen and electron adjusted accurate mass, or neutralaccurate mass.
 55. The method of claim 26, wherein substantiallyequivalent refers ±1 ppm of the hydrogen and electron adjusted accuratemass, or neutral accurate mass.
 56. The method of claim 23, wherein theone or more than one internal control metabolite is aphosphatidylethanolamine (PtdEt).
 57. The method of claim 56, whereinthe phosphatidylethanolamine is selected from the group consisting ofPtdEt 16:0/18:0, PtdEt 16:0/18:1, PtdEt 18:0/18:0, PtdEt 18:0/18:1, andcombinations thereof.
 58. The method of claim 48, wherein the one ormore than one internal control metabolite is a phosphatidylethanolamine(PtdEt).
 59. The method of claim 58, wherein thephosphatidylethanolamine is selected from the group consisting of PtdEt16:0/18:0, PtdEt 16:0/18:1, PtdEt 18:0/18:0, PtdEt 18:0/18:1, andcombinations thereof.